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APPARATUS FOR THE ESTIMATION OF UREA. 
(Seepage sq.) 



CHEMICAL ANALYSIS 



OF 



THE URINE, 



BASED IN PART ON 



(CASSELMANN'S ANALYSE DES HARNS,) 



BY 

EDGAR F. SMITH, Ph.D., JOHN MARSHALL, M.D., 

Asa Packer Professor of Chemistry Demonstrator of Chemistry, Medical De- 

in Muhlenberg College. partment, University of Penna. 



WITH 



ILLUSTRATIONS. 




PHILADELPHIA : 

PRESLEY BLAKISTON, 

1012 WALNUT STREET. 
1881. 

ft 






Entered according to Act of Congress, in the year 1881, by 

PRESLEY BLAKISTON, 

In the Office of the Librarian of Congress at Washington, D. C. 



PREFACE. 



Intimate association with students as instructors in medi- 
cal chemistry has revealed to the authors the fact that 
none of the existing works on urinary analysis deal suffici- 
ently with the chemical side of the subject. Cognizant of 
this, and believing that the requirements of the present 
curriculum demand a more thorough knowledge of details 
than is usually presented, we have endeavored to collect in 
the following pages all matter bearing on the chemical 
analysis of urine which experience has demonstrated to be 
practical and thoroughly reliable. Selecting as a partial 
basis for our work the admirable little publication of Cas- 
selmann — Analyse des Hams — we have added numerous 
methods of analysis and suggestions to enable the student 
at work in the laboratory, or privately, to perform under- 
standingly the solution of the many problems met with in 
the analysis of urine. 

As volumetric methods of analysis are readily applied 
in estimating the urine constituents, the preparation of 
standard solutions and the accompanying calculations have 
received due attention. Following immediately upon the 
close of the chemical portion of the work will be found a 
section upon the microscopic examination of urinary sedi- 

v 



VI PREFACE. 

ments, interesting alike to the student and practitioner of 
medicine. 

The plates illustrating the microscopic character of vari- 
ous urine constituents are borrowed from Casselmann, 
while the apparatus of Huffner, for the estimation of urea, 
is here introduced from the Journal fur prakt. Chemie. Its 
simplicity and accuracy recommend its general adoption. 
For the apparatus pictured in the frontispiece, we are under 
obligations to Prof. Wormley, to whom, as well as to Prof. 
Samuel P. Sadtler, we would here express our sincere 
thanks for the many kindnesses shown us during the pro- 
gress of our labors. S. and M. 



CONTENTS. 



THE URINE. 

Definition of Urine. Urine of Carnivorous Animals. Urine 
of Herbivorous Animals. The Characteristics of these 
two Varieties. The Properties of Normal Human Urine. 
The Normal and Constant Constituents of Human Urine. 
Behavior of Urine with Chemical Reagents. The Acid 
and Alkaline Fermentation of Normal^ Urine. The Re- 
lation of the Acid Fermentation of Urine to the Forma- 
tion of Urinary Deposits and Calculi. The Decomposi- 
tion of the Urea, in the Alkaline Fermentation of Urine, 
into Free Ammonia and Acid Ammonium Carbonate. 
Abnormal, Normal and Accidental Constituents of Urine. 
Apparatus Required for the Examination of Urine. Re- 
agents Necessary in Urinary Analysis 9-14 

II. 

PHYSICAL PROPERTIES AND REACTIONS OF THE URINE. 

Properties of Urine Interesting in Diagnosis. Various Colors 
of Urine. The Conclusions Derived from these Colors. 
The Odor of Human Urine. The Specific Gravity of 
Human Urine. Approximate Determination of the 
Solids in Urine. The Influence of Albumen and Sugar 
on Specific Gravity. Importance of Acid Urine to the 
Practitioner. Formation of Sediments 14-17 

III. 

THE MOST IMPORTANT NORMAL CONSTITUENTS : THEIR OCCURRENCE IN 
NORMAL AND PATHOLOGICAL URINE AND THE CHEMICAL DETECTION 
OF THE SAME. 

The Quantity of Constituents Varies with Manner of Life, 
Nourishment, etc. The Increase and Decrease of Nor- 
mal Constituents. Relation of the Solids and Water in 
Urine. Estimation of the Fixed Salts in Urine. The 
Quantity of Urea in Normal Urine. The Qualitative De- 
tection and Quantitative Estimation of Urea. Occurrence 
of Uric Acid ; its Increase and Decrease. Qualitative De- 
tection of Uric Acid. The Quantitative Estimation of 
Uric Acid. Oxaluric and Hyposulphurous Acids. Chlo- 
rides in Urine ; Decrease in Disease. Qualitative De- 
tection of the Chlorides. Gravimetric Determination of 
the Chlorides. Chlorides Determined Volumetrically. 
Phosphoric Acid in Urine ; its Increase and Decrease. 
Increase of the Alkaline Phosphates. Increase of the 
Phosphates of the Alkaline Earths. Detection and Quan- 
titative Estimation of Phosphoric Acid. Estimation of 
Phosphoric Acid Combined with the Alkaline Earths 

vii 



Vlll CONTENTS. 

(earthy phosphates). Detection and Gravimetric Esti- 
mation of Sulphuric Acid. Volumetric Estimation of 
Sulphuric Acid. Sulpho- Acids Present in Urine. Color- 
ing Matters in Urine. Alterations Suffered by these 
Pigments through Pathological Processes. Detection 
of Urobilin. Approximate Estimation of the Coloring 
Matter. Test for Urophain 17-49 

IV. 

ABNORMAL CONSTITUENTS OF URINE ; THEIR OCCURRENCE AND 
DETECTION. 

Origin of Abnormal Constituents. The Appearance of Al- 
bumen in Urine. Albuminuria and Hematuria. Dis- 
charge of Albumen in Pyuria. Hasmatopyuria. Peptones 
in Urine. Detection of Albumen. Quantitative Estima- 
tion of Albumen. Sugar in Urine. Sugar as a Constant 
Ingredient of Urine. Color of Urine in Presence of Su- 
gar. Qualitative Test for Sugar. Quantitative Determi- 
nation of Sugar. Inosite in Urine ; its Detection. Con- 
ditions Under which Lactic Acid is Noticed in Urine. 
Detection of Lactic Acid. Fats and Fatty Acids. Biliary 
Coloring Matters ; Biliary Salts and Taurin. Occur- 
rence of Leucin, Tyrosin and Cystin. Fibrin in Urine. 
Fibrin Cylinders Under the Microscope. Diseases in 
which the Blood Pigments Occur. Haematin. Occur- 
rence of Blood as such in Urine. Almen's Test for 
Blood in Urine. Hydrogen Sulphide and Ammonium 
Sulphide in Urine. Oxymandelic Acid of Schultzen 
and Riess. Indican ; its Composition and Estimation 49-75 

V. 

URINARY DEPOSITS ( SEDIMENTS). 

Use of the Microscope in the Examination of these De- 
posits. Varieties of Urinary Sediments. Course to be 
Pursued in the Examination of Deposits. Relations of 
Sediments to the Diagnosis of Disease 75-87 

VI. 

PRACTICAL HINTS TO A COURSE FOR THE QUALITATIVE AND QUANTI- 
TATIVE EXAMINATION OF URINE. 

Course to be Pursued in a Practical Examination of Urine. 87-95 

VII. 

URINARY CONCRETIONS. 

Difference Between Gravel and Calculi. Chemical Con- 
stituents of Calculi. Combustible Calculi. Non-com- 
bustible Calculi. Fusible Calculi. Composition of Urine. 
Table for Tension of Aqueous Vapor 95-101 



CHEMICAL ANALYSIS 

OF 

THE URINE. 



THE URINE. 

The urine is that peculiar fluid eliminated by the kid- 
neys, in which we find the elements that have become use- 
less to the animal economy in the form of soluble nitro- 
genous bodies and salts. We can distinguish two varieties 
of urine among the mammals, depending entirely upon 
their nourishment, viz : — 

(a) Urine of Herbivorous animals. 
(6) Urine of Carnivorous animals. 

The first is characterized by its constant cloudy appear- 
ance, its alkaline reaction and the remarkably large quantity 
of phosphates of the alkalies, and alkaline earths present 
in it. Uric acid is entirely absent, while hippuric acid is 
abundant in it. The urine of carnivorous animals in 
a fresh condition is clear, light yellow in color, with an 
agreeable odor, bitter taste (due to urea and indican. K. 
B. Hofmann), and acid reaction. It is rich in urea, but 
almost perfectly free from uric acid, which at the most 
occurs in traces. 

2. Normal human urine resembles the second variety 
b 9 



10 CHEMICAL ANALYSIS OF THE URINE. 

(urine of Carnivorse). Freshly eliminated it is clear, of an 
amber yellow color,with a decided acid reaction (due, accord- 
ing to Liebig, to acid phosphates ; according to Lehmann, 
to free hippuric and lactic acids), and a bitter, saline taste 
and peculiar odor (arising from phenol. Stadeler). Its 
specific gravity varies from 1.005 to 1.025, depending upon 
nourishment, sex, age, etc. 

3. Its normal and constant constituents are water, 
urea, uric acid, hippuric acid, creatin, creatinin, xanthin, 
coloring matters, indican, mucus from the bladder, chlo- 
rides, phosphates and sulphates of potassium, sodium, 
ammonium, calcium and magnesium, and now and then 
traces of iron, nitrates and silica. The gases present are 
nitrogen and carbon dioxide. The quantities of these 
substances are variable, and frequently they occur in such 
minute traces as to render their estimation very difficult. 

4. The behavior of urine with chemical reagents may be 
briefly outlined as follows: On boiling, normal urine should 
remain clear, and generate, when mixed with concentrated 
acids, a peculiar, nauseous odor, and at the same time be- 
come more or less dark in color. Immediate cloudiness 
does not ensue, but in course of time crystals of uric acid 
separate. 

The alkalies precipitate the phosphates of the alkaline 
earths (calcium and magnesium phosphates). 

Barium Chloride in urine acidified with hydrochloric 
acid precipitates sulphuric acid as barium sulphate. 

Silver Nitrate in urine acidulated with nitric acid, throws 
down silver chloride. (If the acidulation be omitted silver 
phosphate will also be precipitated.) 

Ferric Chloride precipitates the phosphoric acid from 
urine previously acidified with acetic acid. 



THE URINE. 11 

Lead Acetate precipitates the chlorides, sulphates and 
phosphates as lead salts. 

Oxalic Acid or Ammonium Oxalate precipitates calcium as 
oxalate. 

Mercuric Nitrate produces in urine, after the removal of 
sulphuric and phosphoric acids, at first a cloudiness, which 
disappears, caused by the following reaction : — 

Hg(N0 3 ) 2 + 2Na CI = Hg Cl 2 -f 2Na N0 3 . 

When this change — the conversion of sodium chloride 
into nitrate, and mercuric nitrate into chloride — is com- 
pleted the further addition of mercuric nitrate will induce 
the separation of a white insoluble compound of mercuric 
oxide and urea. 

Alcohol produces a cloudiness, which disappears upon 
dilution with water. 

5. After protracted standing normal urine undergoes a 
change ; fermentation begins, and this is either — 

(a) Acid fermentation, and afterward 
(6) Alkaline fermentation. 

According to Scherer, the mucus from the bladder con- 
tained in the urine decomposes, forming a fungus very- 
similar to the ferment (Mycodermse Cerevisise), and 
then it decomposes the coloring matter that may be present. 
Usually the color of the urine grows paler in consequence, 
and yields a more acid reaction, due to the formation of 
lactic and acetic acids, and in addition red-colored sedi- 
ments (mixtures of uric acid, urates and mucus) deposit 
out. From this we observe that the acid fermentation of 
urine stands in close relation to the formation of urinary 
deposits and the production of calculi. 

6. Gradually the urine, dependent- on the temperature, 
the cleanliness of the vessels, etc., passes from the acid into 



12 CHEMICAL ANALYSIS OF THE URINE. 

the alkaline fermentation. Indeed, it is not absolutely 
necessary that the acid fermentation should have preceded 
this ; as, under certain circumstances not yet explained, the 
urine enters into the alkaline fermentation in the bladder. 
Here it is induced by the mucous coating of the bladder (ac- 
cording to Tiegheim and Schonbein by distinct, peculiar 
fungi (Torulacese), and it is for this reason that we observe, 
in affections of the mucous membranes of the bladder, that 
the urine that has been recently passed possesses an alka- 
line reaction. 

In the alkaline fermentation of the urine the urea 
decomposes into • acid ammonium carbonate and free 
ammonia — 

CO(NH 2 ) a -f 2H 2 = NH 4 HC0 3 + NH S . 

We notice, in consequence, a strong ammoniacal odor, and 
also that upon the addition of acids to the liquid, strong 
effervescence ensues. The ammonia liberated unites with 
the magnesium phosphate and produces the so-called 
triple phosphates, which separate as a microscopic crystalline 
precipitate. Their form (coffin-lid shape) is characteristic. 
In most cases there is a simultaneous formation of a thin 
coating upon the surface of the urine, and besides, with the 
assistance of a microscope, fungus threads, with and without 
spores, infusoria (vibrionse and monadse), and ammonium 
urate are observed. Mixed with alkalies there follows an 
abundant generation of NH 3 . 

Abnormal Constituents of Urine. 
7. Albumen, glucose, alkapton, inosite, lactic acid and 
lactates, fats and volatile fatty acids, benzoic acid (usually 
converted into hippuric acid), succinic acid, biliary color- 
ing matters, biliary salts, allantoin, leucir, tyrosin, cystin, 






THE URINE. 13 

taurin, mucin, haematin, fibrin, pus, spermatozoids, am- 
monium carbonate, triple phosphate and hydrogen sulphide. 

8. Substances that have been detected in urinary de- 
posits are : uric acid, urates, calcium oxalate and phosphate, 
ammonium magnesium phosphate, ammonium carbonate, 
cystin, tyrosin, xanthin ; and of organized substances : 
mucus and epithelia, pus, blood and spermatozoids, fungi 
and infusoria, fibrin, coagula, sarcinia ventriculi, Goodsir. ■ 

Substances that have received the designation " accident- 
al constituents " are those which, by food or medicine, 
etc., have been introduced into the system and eliminated 
by the urine, partially changed or chemically altered in 
their form. The following have been detected in the urine, 
not altered by their passage through the system : — 

(1) The majority of the salts of the heavy metals, when 
administered in rather large quantities. To this class be- 
long the preparations of antimony, arsenic, mercury, zinc, 
gold, silver, lead, bismuth, etc. 

(2) The alkaline carbonates, potassium iodide, ammoni- 
acal salts. 

(3) The free organic acids. 

(4) The greater portion of the alkaloids. 

(5) The greater portion of the dye and smelling sub- 
stances. 

The following have been found partially x>r entirely 
altered in their chemical nature : benzoic acid, quinic acid, 
cinnamic acid, and oil' of bitter almonds as hippuric acid 
(therefore the occurrence of the latter with Herbivorse) : — 

Tannic acid as gallic acid. 

Alkaline salts of vegetable acids as alkaline carbonates. 

Potassium sulphide as potassium sulphate. 

Free iodine as potassium iodide. 



14 CHEMICAL ANALYSIS OF THE URINE. 

9. Apparatus necessary in the examination of urine : 
Urinometer, a small alcohol lamp or Bunsen gas lamp, a 
water bath, wash bottle, twelve test tubes with stand, fun- 
nels, beaker glasses, porcelain evaporating dishes, watch 
glasses, glass rods, two to four pipettes a 5, 10, 20, and 50 
c.c, a graduated cylinder with foot, filter paper, a polariza- 
tion apparatus for the estimation of sugar, and Vogel's 
color scale. 

10. The reagents that meet with most frequent use are, 
red and blue litmus paper, paper saturated with lead 
acetate, turmeric paper, paper saturated with ammonium 
molybdate, acetic, chromic, hydrochloric, nitric, fuming 
nitric, oxalic and sulphuric acids, ether, absolute and di- 
luted alcohol, distilled water, fused silver nitrate and a so- 
lution of same, barium, calcium and ferric chlorides, Fehl- 
ing's copper solution, fuchsin solution, mercuric nitrate, 
potassium or sodium hydrates, sodium acetate, carbonate, 
nitrate and phosphate, and zinc chloride. 

H. 

PHYSICAL PROPERTIES AND REACTIONS OF URINE. 

11. The physical properties of urine which are of inter- 
est in diagnosis are the color, odor and specific gravity. In 
pathological conditions the normal amber yellow color of 
the urine is converted in some cases into a pale whitish 
yellow, and again to a red or brown black. Hence we dis- 
tinguish as follows : — 

(a) Pale urine — colorless to straw yellow. 

(b) Normal color — gold yellow to amber yellow. 

(c) High colored urine — reddish yellow to red. 

(d) Dark urine — brown, dark beer color to black. 



PHYSICAL PROPERTIES OF URINE. 15 

(e) Green urine. 
(/) Dirty blue urine. 

These different colorations lead us to the following con- 
clusions : — 

(a) The pale urine of patients would suffice to inform 
us that the affected individual was not suffering from any- 
violent, acute, febrile disease. Yet its occurrence may be 
observed in convalescents, and many of those who have 
suffered from some chronic affection (ansemia, chlorosis, 
diabetes). Indeed, if long continued we can determine a 
certain degree of anaemia. 

There are but minute quantities of coloring substances 
and urea in pale urine, and it is generally the case that the 
solid constituents are not abundant except in diabetes 
mellitus, and in healthy persons who drink much water or 
beer (urina potus). 

(b) The normal colored urine justifies the conclusion 
that no sickness is present, in which either the pale urine 
or (c) occur. 

(c) The highly colored urine, by its color and high spe- 
cific gravity r proves conclusively that it is concentrated, 
rich in solid constituents, in urea, etc. The reaction is al- 
ways acid. Persons in good health may, after the inges- 
tion of rich food, eliminate a normal yet highly colored 
urine, but with sick persons the occurrence is of great im- 
portance to the physician, inasmuch as urine of this class 
accompanies all febrile diseases ; in the case of hectic fever 
it forms a more positive guide than the pulse or tempera- 
ture. 

(d) Dark urine generally points to an abnormal pig- 
ment, which is present as an admixture in the urine, e. g., 
biliary coloring matters, coloring matter of the blood, and 



16 CHEMICAL ANALYSIS OF THE URINE. 

also uroxanthin. Not unfrequently the coloration is acci- 
dental, arising from medicaments like rhubarb, senna, car- 
bolic acid and others. 

(e) Green urine of a dirty hue arises from biliverdin, 
in icterus, and brown icteric urine has the same import. 

(/) Dirty bluish urine generally has a dark blue coat- 
ing, and shows a blue deposit formed by the production 
of indigo. The reaction is alkaline. This type of urine 
is met with in cholera and typhus. 

12. The odor of human urine has not yet been referred 
positively to distinct chemical substances. It is merely 
suspected that it is influenced or dependent upon extremely 
minute quantities of phenylic, taurylic, damaluric and da- 
molic acids (Stadeler.) For the practitioner the odor of 
the urine is of but minor significance, as it often varies in 
consequence of the ingestion of foods, medicines, e. g., aspa- 
ragus, oil of turpentine (violet odor), saffron, cubebs, and 
similar substances. 

In alkaline fermentation a disagreeable ammoniacal odor 
is present. Heller observed, in cases of severe typhus and 
spinal troubles, a peculiar musty odor, which indicated the 
formation of fungi (possibly, the cause of the contagious 



13. The changes in specific gravity are worthy of con- 
sideration. The specific gravity of normal urine is greatly 
influenced by the urea and sodium chloride, and can, ac- 
cording to J. Trapp, be employed for an approximate de- 
termination of the solid constituents of the urine. To this 
end ascertain the specific gravity and multiply the two last 
decimal places by 2 (Trapp), or 2.33 (Neubauer). For 
examplej a specimen of urine gave the specific gravity 
1.016; then in 1000 grams there would be about 37 grams 



THE MOST IMPORTANT NORMAL CONSTITUENTS. 17 

of solid matter. Especially important are those cases, 
where in a small volume we find a low specific gravity, and 
in a large volume, a high specific gravity. 

In pathological urine the albumen and sugar most affect 
the specific gravity ; if the latter be high, and the urine 
pale, sugar or albumen would be indicated as present. 

Usually acute inflammations, meningitis, mellituria, in- 
crease the specific gravity, while it is lowered by chronic 
troubles, hydremia and kidney affections. 

14. Normal urine is acid, but can acquire a transitory 
alkalinity through the ingestion of alkaline carbonates, and 
alkaline salts of vegetable acids. Acid urine has some im- 
portance for the practitioner, as it favors the formation of 
certain sediments and concretions and causes an irritation 
of the kidneys and urinary passages (Vogel). The degree 
of acidity of urine increases rapidly in rheumatism, pneu- 
monia and pleuritis. The alkalinity of pathological urine 
should also be carefully noticed. If it originate from po- 
tassium carbonate, it would be one of the most unfavorable 
precursors of brain trouble. Arising from ammonium car- 
bonate, uraemia (the urine often brown colored from ad- 
mixture of hsematin), or catarrh of the bladder (in this 
case, mostly cloudy, from mucus and pus), would very pro- 
bably be indicated. 

m. 

THE MOST IMPORTANT NORMAL CONSTITUENTS : THEIR OC- 
CURRENCE IN NORMAL AND PATHOLOGICAL URINE AND 
THE CHEMICAL DETECTION OF THE SAME. 

15. The normal constituents never occur in any con- 
stant ratio. Their quantity depends : — 

(a) On the manner of life, particularly the nourishment 



18 CHEMICAL ANALYSIS OF THE URINE. 

of the respective individual, his bodily constitution, the 
quality and quantity of nourishment. 

(£>) Upon the time of day and the activity of the excret- 
ing organs. 

(c) Upon the pathological changes. 

A disturbance of the normal proportion of the urine 
constituents is in many instances valuable to the practi- 
tioner in his diagnosis, inasmuch as it has been observed 
that in certain diseases there is not only an increase, but 
also a decrease, of the constituents regarded as normal. It 
is, however, necessary that an accurate knowledge of the 
mode of nourishment, etc., as in a and b, be obtained, 
and in addition, that frequent chemical examinations be 
made. As a consequence of the variation of specific gravity, 
we recognize the fact that the ratio existing between the 
solids and the water in urine cannot be constant; it changes 
from about 12 to 60 grams in 1000 grams of urine. 

16. The solid constituents and water are determined 
quantitatively by evaporating a small and weighed quan- 
tity of the urine upon a water bath, and drying the residue 
in an air bath at 100° C. The method is, however, inac- 
curate, because in the process of drying the acid sodium 
phosphate exerts a decomposing influence upon the urea. 
Therefore, we resort to the use of an apparatus intended to 
catch the ammonia resulting from the decomposition and 
determine it. Or, to avoid any trouble, we determine at 
once the quantity of solids by the specific gravity as given 
§13. 

17. The fixed salts are estimated by evaporating a meas- 
ured volume of urine to dryness and igniting over a naked 
flame until the carbonaceous matter has been completely 
consumed. In doing this care should be taken that (a) the 



THE MOST IMPORTANT NORMAL CONSTITUENTS. 19 

temperature does not become so great as to cause the vola- 
tilization of chlorides, and (b) the carbon does not reduce 
the sulphates and phosphates. To avoid any such risk it is 
advisable before converting the mass entirely into ash, to 
exhaust it with hot water, filter and wash filter paper and 
carbon remaining on it, and the filtrate with the wash water, 
evaporate to dryness, and then heat to a gentle redness 
in a weighed covered porcelain, or better, platinum cruci- 
ble, allow to cool and then weigh. The difference between 
the weight of the empty crucible and the second weight 
will be the weight of the fixed salts. 

18. The quantity of urea occurring in normal urine 
varies, depending largely upon the food ingested and the 
weight of the individual. A mixed diet usually shows 
from 2.5 to 3.2 per cent. 

In all inflammatory diseases, especially in acute brain 
trouble, in rheumatism, and in dropsy, if diuretics be ad- 
ministered the amount of urea is increased. It is decreased, 
on the other hand, by neuralgic processes, chronic diseases, 
wherever a change of the substance underlies the affection, 
in diseases of the spinal cord and kidneys. In typhus, at 
first there is an increase of urea, but it rapidly falls, while 
it rises in meningitis and remains" almost constant in quan- 
tity. 

Qualitative Detection and Quantitative Estimation of Urea. 

19. 20 to 25 c.c. of urine are evaporated to a syrupy 
consistence, upon a water bath, the residue repeatedly ex- 
hausted with alcohol, filtered and the alcohol expelled by 
evaporation upon a water bath. Urea remains behind 
somewhat discolored. (Plate I, Fig. 1.) If it be now dis- 
solved in a small quantity of water, and oxalic or nitric 



20 CHEMICAL ANALYSIS OF THE URINE. 

acid added, combinations of urea with these acids will sepa- 
rate in white shining leaflets or hexagonal plates. (Plate 
I, Fig. 2.) When the urea is present in minute quantity 
the urine is mixed with nitric acid and the formation of 
crystals observed under the microscope. Musculus (Pharm. 
Centralblatt 15, 161) detects urea in solution by means 
of a paper upon which there is a urine ferment. The 
latter is prepared by filtering ammoniacal urine through 
filter paper, washing the filter, drying at 35-40° C, and 
finally the paper is colored with turmeric, again dried and 
preserved in closed glass vessels. This paper retains its 
sensitiveness for some time. To detect urea immerse it in 
a neutral urine, and in the presence of the former it will be 
decomposed by the ferment into ammonium carbonate and 
the paper rapidly becomes brown in various places. 

20. Various quantitative methods for the determination 
of this constituent have been proposed. That of Liebig 
seems to be most generally employed, and yields excellent 
results. On adding a dilute mercuric nitrate solution to a 
dilute urea solution, and neutralizing the free acid gradu- 
ally with sodium carbonate, a voluminous, flocculent pre- 
cipitate will form. Continuing this alternating addition of 
mercuric nitrate and sodium carbonate a moment will occur 
when the solution of mercuric nitrate added will produce, 
with the sodium carbonate, a yellow coloration of mercuric 
oxide or basic mercuric nitrate. The solution will then no 
longer contain any free urea, but this will be in combination 
with mercuric oxide, two equivalents of the latter, 2 HgO 
= 432, to one equivalent of urea CO(NH 2 ) 2 == 60, forming 
2 HgO ; CO(NH 2 ) 2 . For convenience we use a solution of 
HgO in nitric acid, each cubic centimetre of which will 
equal 0.010, or ten milligrams of urea. The reaction 



THE MOST IMPORTANT NORMAL CONSTITUENTS. 21 

occurs between one molecule of urea and two molecules of 
mercuric oxide; and to prepare a standard solution we 
follow the equation — 

60 : 432 : : 10 : x = 72. 

CO(NH a )> : 2HgO : : 10 grms. urea : x = 72 grms. the 
quantity of mercuric oxide to be dissolved in a porcelain 
dish on a water bath in strong nitric acid, and diluted 
with distilled water to 1.000 cubic centimetres. "But ex- 
periment has shown that 5.2 grams HgO should be added, 
to allow for action upon the indicator — sodium carbonate 
— leaving 5.2 milligrams HgO in excess in each cubic cen- 
timetre of the mercury solution over and above the required 
quantity, to unite with the urea. Therefore, dissolve 77.2 
grams HgO,* in strong nitric acid, evaporate excess of 
latter on a water bath until the liquid becomes of a syrupy 
consistence. Treat the residue with water, and dilute to 900 
cubic centimetres.f Knowing the approximate strength 
of the latter solution, we determine its exact titre by 
means of a normal urea solution, prepared by dissolving 
two grams carefully dried urea in a little water, and dilut- 
ing to exactly 100 cubic centimetres with distilled water. 
Then of this solution, 

100 c.c. = 2 grams urea. 
10 c.c. = .200 milligram urea. 
Having done this, we remove 10 c.c. of the urea solution to 
a beaker, and, by means of a burette, gradually add the 
mercuric nitrate solution, mentioned above, until a drop of 
the liquid brought in contact, by means of a glass rod, with 

* Prepared according to Dragendorff, by the precipitation of a solution of 96.855 
grams pure mercuric chloride by dilute sodium bydiate. Wash and dry. 

f In case any basic nitrate of mercury should separate on dilution with water, 
allow it to settle, pour off the supernatant liquid, and dissolve the precipitate in a few 
drops of strong nitric acid, and then add to the original liquid. 



22 CHEMICAL ANALYSIS OF THE URINE. 

a drop of a saturated solution of sodium carbonate, yields 
a yellow precipitate. Note the exact number of cubic 
centimetres of mercuric nitrate used. If the latter solution 
had been exactly standardized, just 20 cubic centimetres 
would be required for the 10 c.c. of the urea solution. The 
number, however, of cubic centimetres of mercuric nitrate 
solution will be less than 20 c.c. Then, in order to bring 
it up to the proper titre, we make the following dilution : 
e. g., if 18.5 c.c. of the approximate mercuric nitrate solu- 
tion were necessary to precipitate 10 c.c. urea solution, 1.5 
c.c. of distilled water must be added for every 18.5 c.c. 
of the original solution, or 15 c.c. for every 185 c.c. of the 
original approximate mercuric nitrate solution. As we 
had at first 900 c.c, and removed 18.5 for experiment, 
there remained 881.5 c.c; then, as the dilution for every 
185 c.c of mercuric solution is 15 c.c, the corresponding 
dilution for 881.5 c.c. would be as many times 15 c.c as 
185 c.c. are contained in 881.5 c.c, or 4.76X15 c.c.= 
71.40 c.c. distilled water, which, when added to the mer- 
curic solution, will bring it up to the proper strength. 

In this method of determining urea in urine, by means 
of mercuric nitrate, it is necessary to remove the phosphoric 
and sulphuric acids from the urine, which is accomplished 
by means of a barium' mixture (1 part of a cold saturated 
solution of barium nitrate, and 2 parts of a cold saturated 
barium hydrate solution). 

Execution of the Method. 

Measure off a definite volume, say 40 c.c. of urine, into 
a beaker glass, add half its volume of the barium mixture, 
then filter through a dry filter, and take 15 c.c from fil- 
trate. These 15 c.c would contain 10 c.c of urine (be- 



THE MOST IMPORTANT NORMAL CONSTITUENTS. 23 

cause the latter had been diluted to half its volume by the 
barium mixture). Now fill a Mohr's burette to the zero 
.mark with the standard mercuric nitrate solution, and 
permit the same to run into this urine mixture, drop by 
drop, until an increase in the precipitate can be no longer 
noticed. Take out a drop from the well-stirred solution, 
by means of a glass rod, place it upon a watch glass and 
bring a drop of the sodium carbonate solution in contact 
with it. If the mixture remains white, continue the addi- 
tion of the mercuric solution to the urine, and repeat the 
test. In this way proceed until the sodium carbonate so- 
lution causes a distinct yellow colored precipitate. The 
number of cubic centimetres of the mercuric solution used 
multiplied by .010 gram will give the number of milli- 
grams of urea contained in the 10 c.c. of urine. This, mul- 
tiplied by 10, will give the quantity in 100 parts, or the 
percentage. 

21. Errors that belong to this method and the correc- 
tions for the same are — 

(a) Corrections for volume of reagent required. 

In standardizing the reagent the proportion by volume 
was 20 c.c. (= 2 vols.) of the reagent to 10 c.c. (1 vol.) of 
the pure urea solution, and as each c.c. of the reagent con- 
tained in excess of that actually required to precipitate 
the urea present 5.2 mgrms. HgO as nitrate, to react 
upon the indicator, the 20 c.c. of reagent employed con- 
tained 5.2 X 20 = 104 mgrms. unprecipitated HgO which 
were finally distributed through 30 c.c. of liquid. Hence 
104 -r- 30 = 3.47 mgrms. of HgO present in each c.c. of the 
final mixture. 

Obviously, a similar proportion will exist when 1 c.c. of 
urine containing 3 per cent, urea are mixed with 5 c.c. 



24 CHEMICAL ANALYSIS OF THE URINE. 

barium mixture, and then 30 c.c. of the mercuric nitrate 
solution added. 

But, when the undiluted urine contains over 3 per cent, 
urea, there will be required for 15 c.c. of the urine mixture 
more than 30 c.c. of the reagent, and consequently the 
excess of HgO present will be under a less degree of 
dilution than existed in standardizing the reagent, and 
therefore the final reading would be a little too low. 

This discrepancy in regard to dilution, when over 30 c.c. 
of the reagent are required for 15 c.c. of the urine mixture, 
may be corrected by adding to the urine mixture £ c.c. of 
distilled water for each c.c. of the reagent employed above 
30, and then repeating the titration. 

Thus, if 40 c.c. of the reagent are employed for the first 
titration, we add to 15 c.c. of the urine mixture 5 c.c. of 
water, and then repeat the titration. 

So, on the other hand, if less than 30 c.c. of the reagent 
are required for 15 c.c. of the urine mixture, the excess of 
HgO present in the reagent will be under a greater degree 
of dilution than was present when the reagent was stand- 
ardized. This difference in conditions may be compensated 
for by deducting .1 c.c. for every 4 c.c. of the reagent re- 
quired less than 30. 

Thus, if 22 c.c. are required — or 8 less than 30 — then 22 
— .2 = 21.8 c.c. the quantity of reagent actually required 
for the precipitation of the urea present, and still leave in 
solution the same relative proportion of HgO to act upon 
the indicator as was present when the reagent was stand- 
ardized. 

(6) In the sodium chloride present. Either remove the 
chlorine with silver nitrate, or if the quantity of sodium 
chloride does not exceed 1 to H per cent., it is only 



THE MOST IMPORTANT NORMAL CONSTITUENTS. 25 

necessary, in order to obtain the approximate number of 
milligrams of urea in 10 c.c. of urine to deduct 2 c.c. 
from the number of cubic centimetres of mercuric nitrate 
required in the estimation. 

(c) When the urine contains albumen remove it before 
the estimation is made, by coagulation and filtration. 

(d) When ammonium carbonate is present add the ba- 
rium mixture, and expel the ammonia by boiling. To esti- 
mate the ammonia titrate the urine with a normal sulphuric 
acid solution. 

Salkowski (Zeitschrift fur physiologische Chemie, 4, 80.) 
asserts that Liebig's method for the estimation of urea 
by means of mercuric nitrate does not yield the quantity 
of urea, but the approximate quantity of nitrogen in the 
urine. From his experiments it appears that in the 
presence of amido and uramido-acids the method furnishes 
not the urea alone, but the entire quantity of nitrogen in 
the liquid. 

Fowler's Method for the Estimation of Urea. 

22. Determine the specific gravity of the urine, and also 
that of a solution of sodium hypochlorite intended to decom- 
pose the urea, then to one volume of the urine add seven 
volumes of hypochlorite solution, multiply the specific 
gravity of the hypochlorite solution by 7, and add the 
result to the specific gravity of the urine. Divide the re- 
sult of the addition by 8, in order to obtain the mean 
specific gravity of the mixture, and in the course of two or 
three hours again determine the specific gravity of the mix- 
ture. Deduct this last specific gravity from the mean spe- 
cific gravity, and multiply the result by .77, and the pro- 
duct will be the percentage of urea. Care must be observed, 



26 CHEMICAL ANALYSIS OF THE URINE. 

in the taking of each specific gravity, that the temperatures 
of the liquids be the same. 

Example : — 
Sp. grav. of urine = 1030 X 1 vol. == 1030 
Sp. gr. of hypochlorite == 1027 X 7 vols. = 7189 



8)8219 



mean sp. grav. = 1027 
and after decomposition the sp. grav. = 1024 

3 X .77 = 
2.31 per cent urea. 

The Hypobromite Method for the Estimation of Urea. 

23. This method is based on the fact that when urea is 
exposed to the action of a hypobromite, decomposition en- 
sues, resulting in the formation of an alkaline bromide, 
carbon dioxide and nitrogen gas. The latter is collected, 
and its volume measured. 
CO(NH 2 ) 2 -|- 3 NaBrO = 3 NaBr + C0 2 + 2 H 2 + N„ 

Urea. Sod. hypobromite. 

Preparation of the Sodium Hypobromite Solution. — The 
directions of Knop should be followed in preparing the 
solution, i. e., dissolve 100 grams sodium hydrate in 250 c.c. 
water, allow to cool, and " mix with it 25 c.c. bromine. 

In making sodium hypobromite two molecules of sodium 
hydrate are required for two atoms of bromine. Knowing 
the density of the latter (about three) we can easily calcu- 
late the approximate quantity of sodium hydrate necessary 
to form hypobromite with the 25 c.c. bromine. Multiply 
the volume (25 c.c.) bromine X 3 (density) = 75 weight of 
bromine. To ascertain how much sodium hydrate will be 



THE MOST IMPORTANT NORMAL CONSTITUENTS. 



27 



required by the bromine we employ the following equa- 
tion: 160: 80 ::75:x 

Br 2 : 2 NaHO : : 75 : x — 37.5 grams, the quan- 
tity of sodium hydrate required by the 75 grams of bro- 
mine, and 100 grams NaHO — 37.5 = 62.5 grams, the 
excess of sodium hydrate which will absorb the liberated 
C0 2 evolved from the urea in the practical use of the 
reagent. 

Execution of the Method. 

HiifFner (Journ. f. prakt. Chemie, Neue. F. Bd. 3, p. 1) 
employs the following apparatus. The vessel -c, of about 
100 c.c. capacity, is in intimate combination with a, of 10-12 
c.c. capacity. They are connected by 
means of a tolerably wide neck (1.5 
centimetres diameter). Between them 
is b, an air-tight glass stop-cock, the 
aperture of which is not more than 7-8 
millimetres wide. The upper contract- 
ed portion d fits closely, by means of 
rubber, the. neck of the upper part of 
the flask that has been prepared for 
the purpose. In this manner there is 
formed a dish, k, of from 4-5 centime- 
tres depth, in the middle of which the 
contracted portion d projects about 1 
centimetre and extends at the same time 
into the eudiometer e, which is about 
30 centimetres long and 2 centimetres' 
wide, divided into | cubic centimetre, and accurately gradu- 
ated. The arms / of the iron stand render the apparatus 
secure. The lower arm clasps the vessel c immediately above 
the cock b, while the upper arm holds e firmly in position. 




28 CHEMICAL ANALYSIS OF THE URINE. 

The urea is determined in this apparatus as follows : 
Aided by a long-necked funnel, fill a and the aperture of 
the stop-cock with the urine, and close the stop-cock. Then 
pour equal volumes of the hypobromite solution and dis- 
tilled water into c, filling it up to the edge. In k pour a 
saturated sodium chloride solution, making a layer 2 centi- 
metres high, which will serve as a bar to the escape of any 
gas. 

During this time a few air bubbles will be liberated 
from c. When they have disappeared, invert over d the 
eudiometer e, filled with water, and when this has been fast- 
ened the preparations cease. With one turn completely 
open the cock b and bring in sudden contact the two solu- 
tions. Owing to its higher specific gravity the hypobro- 
mite solution will sink, mix with the urea solution and in- 
duce the decomposition of the latter with lively evolution 
of nitrogen gas. 

Not more than two or three minutes will elapse from the 
time of the opening of the stop-cock b and the cessation of the 
rapid gas liberation, if the hypobromite solution is concen- 
trated and freshly prepared, and the first contact and mix- 
ture of the solutions has been sufficiently rapid and complete. 
The eudiometer, after standing a while, is carefully removed 
from c, and the volume of nitrogen measured over water, 
as in Dumas' nitrogen estimation. 1 gram of urea, accord- 
ing to its formula, yields 370 c.c. nitrogen at 0° and 760 
mm. pressure. In calculating the result, use the following 
formula : — 

__ 100 v (b — b') . , . , 

P ~ 760. 370. a (1 + 0. 003665 t) m 

p represents the weight of the urea for 100 c.c. urine. 

a represents the volume of urine used. 



THE MOST IMPORTANT NORMAL CONSTITUENTS. 29 

v the volume of nitrogen read off. 

b the barometric pressure. 

t the observed temperature during the measurement of 
nitrogen. 

b' tension of vapor of water for this temperature (see 
Table for Tension of Vapor of Water). 

The urine should be diluted three to four times its volume 
for this method. 

24. The frontispiece represents another very simple and 
convenient form of apparatus, which can be employed in 
the estimation of urea. It consists of a bottle A, contain- 
ing a test tube B, and a large glass cylinder C, in which 
is suspended a graduated burette. The latter is connected 
by means of a rubber tube with A. In making an analysis 
with this apparatus, introduce about 5 c.c. of the urine into 
the test tube B, while about 15 c.c. of the hypobromite solu- 
tion are brought into A, exercising care not to bring the 
liquids in contact with each other. The graduated burette 
is now lowered in C, until the zero mark is on a level with 
the surface of the water in the cylinder, and the connection 
between the burette and the bottle accurately made. A is 
then so inclined that the urine in B will drop into the hypo- 
bromite solution. Decomposition at once occurs, accom- 
panied by effervescence. Gradually raise the burette as the 
nitrogen is evolved, and when the reaction ceases, shake the 
vessel A and allow to stand for a few minutes until it ac- 
quires the temperature of the room in which the operation 
was performed. The water within and without the burette 
is leveled, and the cubic centimetres of nitrogen gas read 
off. This number (say 10 c.c.) multiplied by .027 would 
represent in grams the quantity of urea in 5 c.c. urine. 

For the bottle A can be substituted the apparatus D. 



30 CHEMICAL ANALYSIS OF THE URINE. 

In its arm b introduce with the aid of a pipette a given 
volume of urine, and in c place the solution of hypobromite. 
The connection with the graduated burette is made as 
before. When ready, carefully remove D from the clamp, 
and with the hand slightly incline the vessel, permitting 
the urine to pass drop by drop into the hypobromite 
solution. It is believed that this careful addition of the 
urine to the decomposing agent ensures its complete break- 
ing up. The further manipulations are the same as those 
already described. This piece of apparatus was devised 
by Dr. Williams, of Boston. 

25. It is generally admitted that under the action of the 
hypobromite reagent, and also under that of an alkaline 
hypochlorite, only about 92 per cent, of the total nitrogen 
of the urea is evolved in its free state. M. Mehu, in 1879, 
proposed to remedy this defect by mixing cane or grape 
sugar with the urine, before the addition of the reagent. 

But, quife recently, Professor Wormley has shown that, 
under certain conditions, the whole of the nitrogen is set 
free by the reagent, even without the addition of sugar. 
These conditions, according to this observer, are the folio vy- 
ing :— 

(1) The reagent should be freshly prepared. 

(2) The urea solution should be wholly added to the 
reagent, none of the reagent being allowed to mix with 
the urea solution in the containing bulb or tube. 

(3) The amount of urea operated upon should not ex- 
ceed over one part to about 1200 parts of the somewhat 
diluted reagent. 

It is also important that the urea solution be added to the 
reagent in small portions at a time, thoroughly mixed, and 
the effervescence allowed to cease before any further addition. 



THE MOST IMPORTANT NORMAL CONSTITUENTS. 31 

According to Cotton (Chem. Centralblatt, 1875, p. 263), 
the decomposition of urea by sodium hypobromite is hin- 
dered by certain antiseptics, as sulphurous acid, sulphites, 
hyposulphites, iodine, carbolic acid, etc. ; delayed by such 
as chloral, and hastened by peroxides, acid potassium 
chromate, etc. 

MusculiLs Method for the Estimation of Urea. 

26. Musculns (Archiv.der Physiologie 12, 214), in his in- 
vestigations on urine ferment, remarks that the best material 
for the preparation of the latter is the thick, mucous, 
ammoniacal urine of persons suffering with catarrh of the 
bladder. On adding alcohol to such urine the mucin is 
coagulated to a film-like mass, and can be easily separated 
from the liquid. The precipitate is dried at a gentle heat, 
pulverized and kept in closed glass vessels. 

This ferment is excellently adapted to the quantitative 
estimation of urea. 

10 c.c. of urine mixed with a small quantity of sodium 
carbonate, then diluted 10 times with water, are colored 
with a few drops of litmus, accurately neutralized by a 
dilute acid, 0.2 grams ferment powder added and warmed 
upon a water bath to 35-40° C. In an hour the urea is 
completely decomposed. By titration with normal sul- 
phuric acid the amount of ammonia formed is determined, 
and from this the urea calculated. Creatin and crea- 
tinin are not decomposed by the ferment. 

Uric Acid. 

27. The Uric acid found in urine is partly combined and 
partly uncombined ; its quantity ranges from 0.2 gram to 
1 gram in 24 hours. Disturbed digestion, fevers, affections 



32 CHEMICAL ANALYSIS OF THE URINE. 

of the respiratory organs and disturbance of the blood 
circulation increase the quantity of uric acid, while in its 
decrease it is analogous to urea, and like the latter may be 
converted into ammonium carbonate. 

28. For its detection evaporate, on a water bath, 100 to 
200 c.c. of urine, from which any albumen present has 
been previously removed by coagulation and nitration, to 
a syrupy consistence. Dissolve out the urea and extractive 
matters with alcohol, and the residue will consist of uric 
acid, mucin and fixed salts. 

Add a little nitric acid to a portion of the residue and 
warm, when nearly all will dissolve. On careful evapora- 
tion on a water bath there will remain a red-colored 
residue, which moistened with ammonium hydrate (avoid 
an excess) will assume a purplish-red color — murexide. 
With a drop of sodium or potassium hydrate this becomes 
purplish-blue. 

Another portion of the first residue dissolved in potas- 
sium hydrate, then mixed with hydrochloric acid and 
allowed to stand for some time, will yield crystals of uric 
acid. (Plate i, Fig. 4.) 

When much uric acid is present add hydrochloric acid 
to 200-300 c.c. of urine, and allow the same to stand 
24 hours. In that time the uric acid will have separated 
out in colored crystals, and can be readily recognized under 
the microscope (See Plate i, Fig. 5). 

29. In estimating it quantitatively Ave pursue essentially 
the directions in the preceding section, viz : Mix from 200 
-300 c.c. urine with hydrochloric acid (3-4 c.c), and 
allow to stand for 24-48 hours; the temperature being 
as low as possible. The separated crystals of uric acid 
are collected on a previously washed, dried and weighed 



Plate I. 



Fig. 1. 



Fig. 2. 




Pure urea from an alcoholic solution. 
Fig. 3. 




Hijjpuric Acid from normal human urine. 

Fig. 5. 





Urea Oxalate (upper half); urea nitrate (lower 
half.) 

Fig. 4. 




Various forms of Uric acid from urinary sediments. 

Fig. 6. 



Uric acid. 




NaturaT Sodium Urate. 



THE MOST IMPORTANT NORMAL CONSTITUENTS. 33 

filter paper, washed well with water and after drying, 
weighed. The first weight of the filter paper subtracted 
from the last weight will give the amount of uric acid in 
the quantity of urine employed. 

Salkowski (Virchow's Archiv, 68, 1), proposes the follow- 
ing method for the determination of uric acid : 200 c.c. urine 
are rendered strongly alkaline with sodium carbonate (10 
c.c. of concentrated solution) ; after an hour 20 c.c. of a 
concentrated ammonium chloride solution are added, and 
the whole allowed to stand at a low temperature for 48 
hours, then filtered through a weighed filter and washed two 
or three times with water. The filter is then filled with 
dilute hydrochloric acid (1 part commercial acid to 10 
parts water), and the filtrate preserved. The addition of 
hydrochloric acid to the precipitate on the filter is repeated 
several times, until all the ammonium urate has been con- 
verted into uric acid. Let the filtrate stand six hours ; 
the uric acid that separates from it in this time is brought 
upon the same filter ; wash the precipitate twice with water, 
then with alcohol, until the acid reaction of the filtrate 
passing through disappears, and dry at 110° C, and weigh. 
To the number found add 0.030. Dilute urine should be 
evaporated until its specific gravity becomes 1.017-1.020. 

Oxaluric and Hyposulj)hurous Acids. 

30. Recently Schunk discovered oxaluric acid in normal 
urine, existing there in combination with ammonia. It is a 
white, acid tasting, crystalline powder, difficultly soluble in 
water. The ammonium salt is soluble in water. 

By dissolving uric acid in warm, very dilute nitric acid, 
and adding ammonium hydrate just as the solution is cold, 
then evaporating to crystallization, we can easily obtain 



34 CHEMICAL ANALYSIS OF THE URINE. 

crystals of ammonium oxalurate. Hydrochloric acid 
separates the free oxaluric acid as a white powder from 
concentrated solutions of the ammonium salt. The acid 
dissolved in water and recrystallized forms beautiful aggre- 
gations or rosettes. 

A. Strumpell (Archiv. d. Heilkunde, 17, 390), has dis- 
covered hypo-sulphurous acid in the urine of a typhoid 
patient. The acid can be estimated quantitatively by 
precipitating with barium chloride and in the nitrate 
from the barium sulphate (which will contain barium 
hyposulphite in solution), the barium hyposulphite can 
be oxidized by means of a few drops of nitric acid, and the 
S 2 2 can be calculated from the amount of barium sulphate 
formed. 

The Chlorides in Urine. 

31. These occur principally as sodium chloride. They 
average in 24 hours about 15 grams in 1600-1700 c.c. 
urine. In a healthy, robust man they can become even 
more abundant. 

The decrease of chlorides is of particular interest to the 
diagnostician, and has been noticed : — '■ 

(a) in all cases where the chlorides have not been re- 
absorbed, as in cholera, certain stages of typhus, inanition 
following pathological changes, etc. 

(fi) in abnormal transudations. 

(c) in acute exudations in the following pathological 
processes ; pneumonia, pleuritis, peritonitis, pericarditis, en- 
docarditis, meningitis, typhus, acute miliary tuberculosis, 
and the like. The disappearance of the chlorides in rheu- 
matism of the joints and pericarditis is characteristic. 
Their quantity decreases so rapidly then that by comparison 
of the tests made within a few hours of each other we can 



THE MOST IMPORTANT NORMAL CONSTITUENTS. 35 

determine upon any .conspicuous change in the course of 
the ailment. 

The qualitative test for the detection of the chlorides 
consists in acidifying the urine with nitric acid, then adding 
silver nitrate, when chlorine, if present, will be precipitated 
as silver chloride. 

32. Quantitatively the chlorides can be estimated gravi- 
metrically or volumetrically. 

(a.) A definite volume (say 10 c.c.) of the urine is 
evaporated to dryness on a water bath with a few drops of 
nitric acid and about two grams potassium nitrate. It is 
then ignited over a naked flame until all the carbonaceous 
matter has been destroyed, allowed to cool, dissolved in 
water acidified with nitric acid, heated to almost the boiling 
point, when silver nitrate is added and the solution stirred 
with a glass rod. This will cause the silver chloride to 
collect and settle. The addition of a drop of silver nitrate 
to the supernatant liquid will show whether the precipita- 
tion is complete. If so, filter the solution and bring the 
silver chloride upon a filter, wash rapidly with hot water, 
dry, then separate the precipitate as fully as possible from 
the filter and place it in a weighed porcelain crucible. The 
filter is reduced to ash on the inverted lid of the crucible. 
The traces of silver chloride which are reduced to the me- 
tallic state can be reconverted into chloride by moistening 
the ash with a drop of nitric acid and then a drop of hydro- 
chloric acid. Heat carefully and evaporate excess of acid, 
allow to cool, place the lid upon the crucible, to which 
apply a low heat, then, after cooling, weigh. 

To calculate the quantity of sodium chloride the follow- 
ing equation is employed : — 

AgCl : NaCl : : wt. of prec. : x == amt. of NaCl 
143.5 : 58.5 



36 CHEMICAL ANALYSIS OF THE URINE. 

in the 10 c.c. urine; multiply x by 10 and the percentage 
will be obtained. 

To calculate the quantity of chlorine change the second 
term of the equation to 35.5 and x will equal the quantity 
of that element in a given quantity of the urine. 

(b.) Of the volumetric methods there are several. 

I. That of Liebig is based on the circumstance that 
sodium chloride acting upon mercuric nitrate causes the 
formation of the soluble compounds, mercuric chloride and 
sodium nitrate, and that so long as there is a chloride 
present in the urine a precipitation of the urea cannot occur, 
but just as soon as all the chlorine has entered into combi- 
nation with the mercury and the mercuric nitrate is added 
in more than sufficient quantity to cause the preceding 
change with the chloride, a white cloudiness will appear, 
resulting from the union of the excess of mercuric oxide 
with urea. This latter substance then acts as the indicator, 
and on this behavior the method is founded. As one 
equivalent of mercuric oxide is equal to two of sodium 
chloride, the solution of mercuric nitrate is so prepared that 

1 c.c. of the latter will equal ten milligrams of sodium 
chloride. That is, we would make the following calculation 

HgO = 2 NaCl and 
117 : 216 :: 10 : x 

2 NaCl : HgO : : 10 grams NaCl : x == 18.461 grams 
of mercuric oxide, which are to be dissolved in a porcelain 
dish, on a water bath, with strong nitric acid, and treated in 
the same manner as described in the preparation of the 
mercuric nitrate solution for the estimation of urea, (page 
21). After dilution it can then be standardized by means 
of a standard solution of sodium chloride, prepared by dis- 
solving one gram perfectly dried sodium chloride in 100 



THE MOST IMPORTANT NORMAL CONSTITUENTS. 37 

c.c. of water. 10 c.c. of the latter solution are measured out 
iuto a beaker, a small pinch of urea dissolved in it and the 
mercuric solution added until the appearance of a perma- 
nent cloudiness. The quantity used is then read off. If, 
for example, 8.2 c.c. mercuric solution were required, then 
for every 8.2 c.c. of mercuric solution on hand add 1.8 c.c. 
distilled water. A new titration can then be made and the 
solution will be found up to the proper strength. 10 c.c. 
should equal 10 c.c. of the sodium chloride solution. 1 c.c. 
of the mercuric solution will then be equal to 10 milli- 
grams sodium chloride or 0.00606 milligram chlorine.* 

In the practical execution of this method the phosphoric 
and sulphuric acids must first be removed from the urine, 
which is accomplished by the use of the barium mixture, 
as given in the determination of urea. Take 20 c.c. of 
urine, mix with 10 c.c. barium mixture, and filter through 
a dry filter. The solution will be alkaline ; and from 
the filtrate take 15 c.c. (of which 10 c.c. are urine) and 
make it neutral, or, at the most, very slightly acid, 
with nitric acid, and then commence the addition of the 
mercuric nitrate, drop by drop, from a burette. The first 
drop of the latter will cause a turbidity, which disappears 
on stirring the liquid. Continue adding the mercuric solu- 
tion until a permanent turbidity is produced ; read off the 
number of cubic centimetres of the mercuric solution used 
and multiply these by .010, and the product will represent 
the number of milligrams of sodium chloride contained 
in the 10 c.c. of urine. This product multiplied by 10 
will give the percentage of sodium chloride. 

II. Neubauer's modification of Mohr's method. 

10 c.c. urine are brought into a platinum or porcelain 

* NaCl CI NaCI 

58.5 : 35.5 : : .010 : x = 0.0060(5 



38 CHEMICAL ANALYSIS OF THE URINE. 

dish, 2 grams powdered potassium nitrate, free of chlo- 
rine, added, and the whole evaporated to dryness on a 
water bath or hot plate. The residue is heated gently at 
first, over a naked flame, more intensely later, until the 
carbonaceous matter is completely oxidized, and the fused 
mass, upon cooling, is perfectly white in appearance. In 
cooling, withdraw the flame slowly, so as to prevent any 
likelihood of cracking the porcelain dish or spurting of the 
fused substance. The residue of salts is now dissolved in 
about 30 c.c. of water, washed into a beaker, the platinum 
or porcelain dish carefully washed out, and the wash water 
added to the solution of the salts, and the whole evapor- 
ated down to about 30 c.c. The solution will be alkaline, 
from the decomposition of the potassium nitrate. Dilute 
nitric acid (or acetic, in which case neutralization with cal- 
cium carbonate is unnecessary, as this acid does not decom- 
pose silver chromate) is added, drop by drop, to the liquid, 
until the latter yields a faint acid reaction, which is re- 
moved by the addition of a small quantity of precipitated 
calcium carbonate. The latter, if added in excess, need 
not be filtered off. To the solution thus prepared add 2 to 
3 drops of a cold saturated solution of neutral potassium 
chromate, K 2 Cr0 4 , which acts as the indicator. Silver 
has a greater affinity for chlorine than for chromic acid ; 
therefore, no combination will take place between the silver 
and chromic acid until all the chlorine has been satisfied. 
The standard silver nitrate solution is now allowed to run 
into the sodium chloride solution, drop by drop, from a 
burette, with constant stirring, until a distinct orange color 
is produced, which remains permanent*. The number of c.c. 

* Correction. — On account of the dilution of the mixture if more than 10 c.c. of the 
silver solution be required to produce the orange coloration, -^ of a c.c. is deducted 
from the number of c c. silver solution for every 5 c.c. used above 10 c.c. 



THE MOST IMPORTANT NORMAL CONSTITUENTS. 39 

silver solution used, multiplied by .010 gram, will give the 
number of milligrams of sodium chloride in the 10 c.c. em- 
ployed. This number multiplied by 10 will furnish the 
percentage. The amount of chlorine is found by multiply- 
ing the number of c.c. silver solution used by 0.00606 gram, 
and the product multiplied by 10 gives the percentage of 
chlorine." 

Preparation of Silver Nitrate Solution. 

The standard silver nitrate solution is prepared as fol- 
lows : It is made of such strength that 1 c.c. will be equal 
to 10 milligrams sodium chloride. The reaction takes 
place between one molecule of silver nitrate and one mole- 
cule of sodium chloride, represented by the equation — 

AgN0 3 + NaCl = AgCl + NaN0 3 . 
Therefore, in order to determine the quantity of silver ni- 
trate necessary to make a solution of standard strength, we 
use the following proportion : — 

58.5 : 170 : : 10 : x. 

NaCl : AgN0 3 : : 10 grms. NaCl : 29.059 grms. AgN0 3 . 
That is, 29.059 grams chemically pure, fused silver nitrate 
are dissolved in a little water, and the solution diluted to 
1 litre ; then 

1000 c.c. = 10 grms. NaCl. 
1 c.c. = .010 grm. NaCl. 
If necessary, the solution can be standardized by means of 
a standard solution of sodium chloride, made as follows : 
dissolve one gram thoroughly dried sodium chloride, in a 
little water, and dilute to 100 c.c. with distilled water ; 
measure off into a beaker 10 c.c. of the solution, add two or 
three drops of potassium chromate solution, run in from a 
burette, with constant stirring of the liquid in the beaker, 
the silver solution, until an orange coloration appears, 



40 CHEMICAL ANALYSIS OF THE URINE. 

which is persistent. 10 c.c. of the silver solution should 
have been required to produce this coloration, and if more 
or less than this quantity were required, then make the 
necessary corrections, as given under Liebig's mercuric ni- 
trate solution for the estimation of urea. 

III. Primbram's Method. Primbram has proposed a 
slight modification to Neubauer's method. Instead of de- 
stroying the organic matter by ignition with an alkaline 
nitrate, he adds to a measured volume of urine a few c.c. of 
a saturated solution of potassium permanganate, and heats 
to almost boiling, when oxide of manganese separates. 
The permanganate is added until the liquid, on warming, 
retains a purple color, when it is filtered, the precipitate 
washed with hot water, and the colored filtrate decolorized 
by the addition of a little oxalic acid. Any excess of the 
latter is neutralized by a little precipitated calcium carbo- 
nate. The solution is now reduced by evaporation to a 
definite volume (say 10 or 20 c.c, or the original volume 
employed), and titred with the silver solution, as in Mohr's 
method. 

IV. Falck (Berichte d. deutsch. Chem. Gesellschaft, 8, 
12) recommends the following in estimating chlorides in 
urine : after the evaporation of 10 c.c. urine, and ignition of 
the residue with potassium nitrate, the salts remaining are 
dissolved in a little water, and washed into a beaker glass ; 
the alkaline solution is acidified with nitric acid, and, after 
the addition of 4 c.c. ammonium ferric sulphate solution, 
made blood-red by the aid of 1 or 2 drops of a titrated 
ammonium sulphocyanide solution. The standardized silver 
nitrate solution is now added from a burette, until the red 
coloration just disappears. The number of cubic centi- 
metres of the latter solution thus required do not exactly 



THE MOST IMPORTANT NORMAL CONSTITUENTS. 41 

correspond to the chlorine in the liquid, because, in the in- 
cineration with potassium nitrate, nitrites are invariably 
produced, and the nitrous acid liberated upon the addition 
of nitric acid affects the final reaction. Therefore, again 
ignite 10 c.c. urine, strongly acidify the solution with nitric 
acid, mix the solution with an excess of silver nitrate solu- 
tion, so that all the chlorine present will be in combination 
with the silver. The solution is now warmed upon a water 
bath, to expel the nitrous acid, then cooled, mixed with 5 
c.c. iron alum solution and the ammonium sulphocyanide 
added, drop by drop, until the red coloration of iron sulpho- 
cyanide no longer disappears. The difference between the 
required number of cubic centimetres of silver and sulpho- 
cyanide solutions represents the chlorine contained in the 
urine. 

The following solutions are necessary in the above 
method : (a) Solution of silver nitrate, of which 1 c.c. cor- 
responds to 10 milligrams sodium chloride. 

(6) Solution of ammonium sulphocyanide accurately stan- 
dardized with the silver solution, so that 10 c.c. of the 
former will be required to precipitate the silver in 10 c.c. 
of the standard silver solution as silver sulphocyanide. 

(c) A cold saturated solution of crystallized ammonium 
ferric sulphate free from chlorine. 

PHOSPHORIC ACID. 

33. The phosphoric acid in urine exists partly combined 
with sodium, as acid sodium phosphate, and partly in com- 
bination with calcium and magnesium, as calcium and 
magnesium phosphates. Regarding the increase or de- 
crease of phosphates in pathological changes the following 
may be observed : — 

(a) In the urine of persons suffering from inflammatory 

D 



42 CHEMICAL ANALYSIS OF THE URINE. 

diseases, e. g., acute brain affections, acute spinal troubles, 
the alkaline phosphates are increased. They decrease in 
neurosis, chronic spinal affections, and kidney diseases. 

(6) The phosphates of the alkaline earths (earthy phos- 
phates) are increased by meningitis, especially in acute 
brain affections and rheumatism. They decrease in kidney 
and spinal affections, and in neuralgia. 

Detection and Quantitative Estimation of Phosphoric Acid. 

34. On adding ammonium hydrate in excess to urine the 
phosphates of calcium and magnesium are precipitated ; 
the latter as triple phosphate. The phosphoric acid that 
yet remains in solution, after adding ammonium hydrate, 
is recognized by acidifying the solution with acetic acid, 
and then adding a little ferric chloride, when a yellowish- 
white precipitate of ferric phosphate is produced. 

35. Phosphoric acid is best determined quantitatively, 
volumetrically, by means of a standard uranium acetate 
solution. The method is based on the insolubility of ura- 
nium phosphate in acetic acid. The merest trace in excess 
of uranium acetate is recognized by the reddish-brown color 
formed when a drop of the liquid is brought in contact with 
ferro cyanide of potassium. 

The uranium acetate solution is so standardized that 
1 c.c. of it equals 0.005 gram of phosphoric acid. 

The formula of the precipitate formed by the addition of 
the uranium acetate to a solution of a phosphate is 2Ur0 3 : 
P 2 5 +Aq. Two molecules, Ur0 3 are required to combine 
with one molecule, P 2 5 , therefore, in the preparation of 
the standard uranium acetate solution we use the following 
proportions : — 



THE MOST IMPORTANT NORMAL CONSTITUENTS. 43 

142 : 576 : : 5 : x 

P 2 5 : 2Ur0 3 : : 5 grams P 2 5 : x = 20.28 grams 
Ur0 3 , necessary to combine with 5 grams P 2 5 ; then to 
find the quantity of uranium acetate equivalent to 20.28 
grams Ur0 3 we make another proportion : — 

288 : 442 : : 20.28 : x 

Ur0 3 : Ur0 3 (C 2 H 3 2 ) 2 + 2H 2 : : 20.28 grams Ur 
3 : x = 31.1 grams uranium acetate, to be dissolved in 
900 cubic centimetres of water, about 5 c.c. strong acetic 
acid added and allowed to stand for a few hours, in order 
that a precipitate which usually forms may subside. The 
solution is then filtered and titrated by means of a standard 
phosphoric acid solution, and diluted after the plan used in 
standardizing the mercuric nitrate solution for the estima- 
tion of urea (page 22). 

If uranium nitrate be preferred in the preparation of 
the uranium solution, substitute in the second equation 
above, the molecular weight of uranium nitrate (Ur0 3 N 2 
5 -J- 6H 2 = 504) for that of uranium acetate Ur0 3 
(C 2 H 3 2 ) 2 -f 2H 2 = 442), and the result will be the num- 
ber of grams uranium nitrate required. 

Instead of ascertaining the amount of uranium acetate or 
nitrate by the two equations above mentioned, we can 
immediately determine the required quantity of the respec- 
tive compounds by the following single equations : — 

142 : 884 : : 5 : x 

P 2 5 : 2Ur0 3 (C 2 H 3 2 ) 2 + 2H 2 : : 5 grams P 2 5 : x = 
31.1 grams uranium acetate. 

142 : 1008 : : 5 : x 

P 2 5 : 2Ur0 3 N 2 5 + 6H 2 : : 5 grams P 2 5 : x =± 
35.5 grams uranium nitrate. 

The solution of phosphoric acid which is used for 



44 CHEMICAL ANALYSIS OF THE URINE. 

standardizing the uranium solution is prepared as follows : 
As phosphoric acid itself cannot be weighed, a stable 
weighable compound in which it exists in combination with 
a base is used for the purpose. Sodium hydrogen phos- 
phate is the salt usually employed. To obtain one mole- 
cule of P 2 5 we must use two molecules of Na 2 HP0 4 -f- 12 
H 2 0. Then to find the quantity of sodium hydrogen phos- 
phate which shall contain 5 grams phosphoric acid, we use 
the following proportion : — 

142 : 716 : : 5 : x 

P 2 5 : 2Na 2 HP0 4 + 12H 2 : : 5 grams P 2 5 : x = 
25.211 grams sodium hydrogen phosphate, which will be 
equal to 5 grams P 2 5 . The 25.211 grams well crystal- 
lized Na 2 HP0 4 -j- 12H 2 are dissolved in a little water 
and the solution diluted to 1 litre. Then of this solu- 
tion — 

1000 c.c. = 5. grams P 2 5 . 
1 c.c. = 0.005 gram P 2 5 . 

In standardizing, measure off into a beaker 20 c.c. of 
the standard sodium phosphate solution, add 30 c.c. dis- 
tilled water and 5 c.c. sodium acetate solution (prepared 
by dissolving 100 grams crystallized sodium acetate in 900 
c.c. water, and adding acetic acid until the volume reaches 
1000 c.c). The mixture is then heated on a water bath, 
to a temperature between 90 and 100° C, and the uranium 
solution gradually added from a burette, the mixture being 
stirred constantly, until a drop of the liquid, removed by 
means of a glass rod, produces a reddish-brown color 
when brought in contact with some powdered potassium 
ferrocyanide or a concentrated solution of the same. 
20 c.c. of the uranium solution equal to .100 gram P 2 5 



THE MOST IMPORTANT NORMAL CONSTITUENTS. 45 

should be required to unite with the P 2 5 present and give 
the reaction with the indicator — potassium ferrocyanide. 

In this titration a less number of cubic c.c. than 20 
of uranium solution will be used, and then, in order to 
bring it to the exact strength, that is, that 20 c.c. shall be 
required, make the dilution as given under the standard- 
izing of the mercuric nitrate solution. For example, if 
18.4 c.c. uranium solution had been required, then for 
'every 18.4 c.c. contained in the original volume of uranium 
solution add 1.6 c.c. distilled water. 

In the actual analysis, measure off 50 c.c. urine into a 
beaker, add 5 c.c. sodium acetate solution and heat upon 
the water bath. Then slowly add the uranium solution, 
from a burette, with constant stirring of the mixture, until 
a drop of the latter, removed with the aid of a glass rod, 
gives a perceptible reddish-brown coloration when brought 
in contact with the indicator, potassium ferrocyanide. The 
number of cubic centimetres of uranium solution used is 
now read off, and then multiplied by the strength of 1 c.c. 
= .005 gram, and the result will be the quantity of P2O5 
in the 50 c.c. urine employed. 

36. To estimate the phosphoric acid combined with the 
alkaline earths, add to a measured quantity of urine (say 
200 c.c.) ammonium hydrate in excess, and stand aside for 
a few hours, collect the precipitate of earthy phosphates on 
a filter, wash and dissolve in as little acetic acid as possible, 
dilute the solution with water to 50 c.c, add 5 c.c. acetate 
of sodium solution, warm on water bath, and titrate with 
standard uranium acetate solution. The number of cubic 
centimeters of the uranium solution, multiplied by the 
strength of 1 c.c. (0.005 gram), will furnish the quantity 
of P 2 5 combined as earthy phosphates in 200 c.c. urine. 



46 CHEMICAL ANALYSIS OF THE URINE. 

In this determination care should be taken to avoid an 
excess of sodium acetate, as it affects the delicacy of the 
reaction of potassium ferroeyanide. 

SULPHURIC ACID. 

37. Next in importance to the phosphates are the sul- 
phates, which are qualitatively detected in acidified urine 
by means of barium chloride. (See § 4.) 

The quantitative determination of the sulphuric acid is 
based on the insolubility of barium sulphate. The method 
may be executed gravimetrically or volumetrically. If the 
latter is preferable, a standard solution of barium chloride 
should be used ; 1 c.c. of this solution should correspond to 
0.010 gram of sulphuric acid. 

To estimate it gravimetrically add about 20 grams po- 
tassium nitrate to 100 c.c. of urine, and evaporate to dry- 
ness on a water bath, then incinerate over a naked flame 
until all the carbonaceous matter has been destroyed. The 
fused mass is then dissolved in water, acidified with hydro- 
chloric acid, brought to the boiling point and an excess of 
barium chloride solution added. The precipitated barium 
sulphate is collected on a filter, washed with hot water, 
dried, as much of it as possible detached from the filter 
paper and placed in a weighed crucible, the filter paper 
incinerated on the end of a platinum wire, held over the 
crucible, and after cooling, the whole weighed. The fol- 
lowing equation will serve for the calculation of the 
result : — 

233 : 80 : : : x 

BaS0 4 : S0 3 : : wt. of prec. : x 

In addition to the sulphates in urine, Baumann (Be- 
richte d. deutsch. Chem. Gesell, 9, 54,) has proven that 



THE MOST IMPORTANT NORMAL CONSTITUENTS. 47 

sulpho-acidsare also constantly present. According to this 
chemist, the phenol, indigo, and brenz-catechin forming 
substances are found in urine as sulpho-acids. To estimate 
them when both are present, pursue the following course: 
Strongly acidify the fresh urine with acetic acid, and add 
an excess of barium chloride, filter off the precipitate after 
standing one to two hours, wash first with water, then with 
warm dilute hydrochloric acid, and finally with water. 
The filtrate and wash water from the precipitate are then 
warmed for several hours with an equal volume of hydro- 
chloric acid upon a water bath. The precipitate that sepa- 
rates contains, in addition to an amorphous organic sub- 
stance, barium sulphate, the sulphuric acid of which did 
not exist as sulphate in the original urine. 

COLORING MATTERS IN URINE. 

38. Urine Brown, urophain, increases in inflammatory 
troubles, in disorders of the liver, and in icterus, frequently 
very markedly decreased in neurosis. 

Urine Yellow, uroxanthin, is increased in violent func- 
tional disturbances in the spinal marrow (forming urrho- 
din and uroglaucin), e. g., in a sudden fall, sudden fright, 
etc., in acute kidney affections, and in cholera. 

Urine abundant in uroxanthin deposits upon long stand- 
ing, and during alkaline fermentation, a blue sediment 
(uroglaucin) ; hence the so-called blue urine (Cholera mor- 
bus Brightii). 

Urobilin might also be noticed. Jaffe noticed this in 
both normal and pathological urine, and also in the bile. 
The pigment is distinguished by the magnificent fluores- 
cence which it exhibits under certain conditions, and by its 
characteristic spectrum. The urine of persons suffering 



48 CHEMICAL ANALYSIS OF THE URINE. 

with fever is rich in this pigment. The spectroscopic study 
of it reveals an absorption band between Frauenhofer's lines 
b and F. With alkalies it shows a characteristic play of 
colors. 

To detect urobilin in normal urine precipitate 100 to 
200 c.c. of the latter with lead acetate, and decompose the 
washed and dried precipitate with an alcoholic solution of 
oxalic acid. If the solution does not exhibit any absorp- 
tion lines, mix it with chloroform, and shake up with water. 
Upon the addition of ammonium hydrate and zinc chlo- 
ride, the acid alcoholic liquid will yield an exquisite fluo- 
rescence, and show sharp, well defined lines in the spec- 
trum. 

APPROXIMATE ESTIMATION OF THE COLORING MATTERS. 

39. For this purpose either Neubauer and Vogel's color 
scale or Heller's urophain reaction is employed. In 
making the latter we proceed as follows : Pour 2 c.c. of col- 
orless sulphuric acid into a small beaker and let flow into 
this, from a height of about four inches, two parts urine, in a 
delicate stream. The urine, when mixed intimately with 
the sulphuric acid, produces an intense garnet-red colora- 
tion, providing the sample was normal urine, i. e., having a 
specific gravity of 1.020, and the quantity eliminated in 
twenty-four hours being about 1500 c.c. If there has been 
an increase in the quantity of coloring matter, the urine 
mixture will be opaque and black ; if the quantity be less 
than normal the mixture will appear pale garnet-red and 
perfectly transparent. 

Care must be observed in this experiment, that the urine 
does not contain any sugar, blood, or biliary coloring mat- 



T HE ABNORMAL CONSTITUENTS OF URINE. 49 

ter, as these would indicate an apparent increase of the 
quantity of urophain. 

To perform the test for urophain, pour about 3-4 c.c. of 
pure concentrated hydrochloric acid into a small beaker, 
and then drop in, while stirring, from ten to twenty drops of 
normal urine. Usually the quantity of this coloring matter 
is so slight under normal conditions that the acidulated 
urine is of a feeble yellowish-red color. When the quantity 
is large the hydrochloric acid is colored from violet to blue. 
Frequently 1-2 drops of urine suffice to color 4 c.c. hydro- 
chloric acid blue. If a violet color does not appear in from 
one to two minutes, the coloring substance has not increased 
above normal, even if the mixture, after standing from ten 
to fifteen minutes, assumes a dark reddish-brown color. In 
icteric urine the bile-coloring matters should be removed 
with lead acetate and the filtrate employed for this test. 

IV. 

THE ABNORMAL CONSTITUENTS OF URINE; THEIR 
OCCURRENCE AND DETECTION. 

40. These arise in certain disturbances of the health of 
an individual, and are partly such substances which pass 
through the kidneys in consequence of altered transudation 
relations, while they are constantly present in the blood ; 
or they arise from a metamorphosis of the tissues, and are 
even formed in the latter, and under normal conditions 
even further transposed, and under abnormal conditions 
passing through the blood are eliminated by the kidneys. 

ALBUMEN. 

41. The conditions under which albumen appears in the 



50 CHEMICAL ANALYSIS OF THE URINE. 

urine are by far more numerous than formerly supposed, 
when it was believed that from the presence of albumen 
certain diseases could be diagnosed. 

Albumen is found 

(a) In general sickness, e. g., pure hydremia, chlorosis, 
endemic diseases, dropsy. Further, in disturbance of the 
circulatory organs, heart troubles, and affections of the 
liver, when, by a difference of pressure, there ensues an in- 
filtration of albumen. 

(b.) In diseases of the uropoetic system, e. g., in the so- 
called sympathetic kidney diseases, in typhus, peritonitis and 
violent phlogosis which influence hyperemia of the kidneys. 
And in the so-called idiopathic affections of the kidneys : 

(1) Albuminuria such as is observed in Bright's disease, 
in nephritis, neoplasma renis. The so-called Bellinic 
casts, pus sediment in acid reaction and a small quantity of 
neoplasms even, always distinguish each of these troubles 
introducing albumen. 

(2) Hematuria, which may be partly a hemorrhagic 
capillary hematuria, in which fibrous coagula do not ap- 
pear ; or partly a hemorrhagic vascular hematuria, in which 
a blood clot is found ; thirdly, and finally, a serous hema- 
turia, where no blood corpuscles, but blood coloring matters 
are present, together Avith the albumen. If these occur 
where the specific gravity is over 1.020 it is a symptom of 
uraemia. 

Urine, red in color, rich in albumen, free of blood cor- 
puscles, and having a specific gravity below 1.020 is sup- 
posed to contain blotches or collections of blood cells. 
When the specific gravity rises above 1.020 the quantity of 
the blood coloring matter in the urine can only be ac- 
counted for by the ammonium carbonate, which extracts 



THE ABNORMAL CONSTITUENTS OF URINE. 51 

the haematin and becomes thereby a specific ursemic 
symptom. 

42. Very often in pyuria albumen is discharged regu- 
larly. The acid or renal pyuria we find in pyelitis, ure- 
thral catarrh, nephritis, etc. The alkaline pyuria shows 
catarrh of the bladder, in combination with renal pyuria 
or alone, when, however, it is in the pus stage. 

43. Finally, when with the albumen, which passes off 
with pus in phlogosis of the kidneys, not unfrequently an 
equal or greater quantity of albumen in the interstitial 
capillary or vascular hematuria is separated in the urine. 
We designate this stage hsematopyuria.* 

44. We find albumen in urine, in addition, in many fevers, 
remittent as well as intermittent; also in exanthematous 
affections (measles, scarlet fever, smallpox), further in affec- 
tions of the. respiratory organs (pneumonia, tuberculosis), 
and after excesses in eating, and after excitement of the 
animal passions; also after the inhalation of hydrogen 
arsenide. 

Gerhardt (Wien med. Presse, 1871, p. 1.) has frequently 
observed peptones in urine free of albumen,, either as a 
forerunner or consequence of ordinary albuminuria. Sena- 
tor declares that peptones exist in every albuminous urine 
in slight quantities. 

Detection of Albumen. 

45. Many difficulties are met with when testing for albu- 
men. The first step should be to ascertain the reaction of 
the urine. Then, if it be neutral or alkaline, acidulate it 
slightly with nitric acid, and heat the specimen, in a test 
tube, to 60 or 80° C. Turbidity follows, and very soon re- 

* See Folwarczny's Handbuch d. physiolog. Chemie, Wien, 1863. 



52 CHEMICAL ANALYSIS OF THE URINE, 

suits in the coagulation of the albumen. Alcohol also pro- 
duces coagulation. 

Heller's test is to take a small beaker or large test tube, 
bring into it about 10 c.c. of urine, then incline the glass, 
let half this volume of concentrated nitric acid trickle 
down the side, and at the point of contact of the two 
liquids, if albumen is present, there will be produced a 
band-like, sharply denned white zone. It is true that in the 
presence of large quantities of urates in the urine a similar 
layer is produced, not at the point of contact of the urine 
and acid, but higher up, and it is not sharply denned below, 
but is rolled up similar to rising smoke. The other methods 
of estimating the albumen by alcohol and tannic acid we 
will pass over. 

Galipe (Pharm. Zeitschrift fiir Russland, 13, 683) recom- 
mends the following test for albumen in urine. In using 
it the mistaking of phosphates and urates for albumen is 
impossible. Fill a reagent glass one- third with a highly- 
colored picric acid solution, and drop in two to three drops 
of the urine under examination. In the presence of albu- 
men there forms immediately a sharply denned white tur- 
bidity. On warming the liquid the albumen collects into 
balls, which rise to the surface of the liquid and float there. 

Quantitative Estimation of Albumen. 

46. The urine is first filtered, and from 20 to 100 c.c. of 
the filtrate are then taken for the estimation of the albumen 
(we should never have more than from 0.2 to 0.3 gram 
coagulated albumen). Concentrated urine — that is, urine 
containing a large percentage of albumen, should be dilu- 
ted with water. The beaker containing the urine under 
examination is heated on a water bath for half an hour. 



THE ABNORMAL CONSTITUENTS OF URINE. 53 

If a flocculent precipitate does not appear, from want of 
sufficient acidity of the urine, add, by means of a pipette, 
from one to three drops acetic acid, avoiding an excess. 
When the coagulation is complete, filter through a pre- 
viously dried and weighed filter. When the .liquid has 
passed through, wash the albumen with hot water, until a 
drop of the filtrate evaporated on platinum foil does not 
leave a residue. The filter and precipitate are dried on 
a watch crystal at 100° C, and when cooled weighed. 
After the deduction of the weight of the watch crystal and 
filter paper, we have the weight of the albumen. One 
source of error in this method is that in the coagulation of 
the albumen it may enclose earthy phosphates, and there- 
fore, after ascertaining the weight of albumen, place the 
latter in a weighed crucible and ignite, allow to cool, weigh, 
deduct the weight of the crucible -f- the earthy phosphates 
from the first weight (crucible -j- albumen) and the result 
will be the exact amount of albumen. 

The method of Bornhardt for the estimation of albumen 
is readily applied and consequently of service to the prac- 
titioner. It consists in determining the sp. grav. of the 
freshly precipitated albumen. A delicate sp. grav. bottle 
is filled with water and weighed, then the moist albumen 
introduced and the sp. grav. bottle re-weighed. The albu- 
men being specifically heavier than water (1.314), the sp. 
grav. bottle would, of course, show an increased weight in 
the second weighing. The quantity of albumen is found 
from the following formula: — 

x = d, 1.314 



0.314, 
in which d represents the difference in weight of the specific 



54 CHEMICAL ANALYSIS OF THE URINE. 

gravity bottle when filled only with water, and then with 
water and albumen. 

SUGAR IN URINE. 

47. Brficke contended that sugar in small quantities was 
a normal constituent of urine, but this view has not met 
with general acceptance. It is a constant ingredient, how- 
ever, of urine in but one disease — diabetes mellitus. Here 
it is eliminated, frequently in such abundance that the 
urine possesses a sweet taste, and cloths soaked in it, after 
the volatilization of the urine, become sticky, and look as if 
they had been coated with honey. Sugar appears in the 
urine after injury to the fourth ventricle of the brain ; 
therefore, this was believed to be the cause of the disease in 
diabetes mellitus, but the connection between the irritation 
of the brain and the sugar separation is yet perfectly in 
the dark. Sugar is also found in galactostasis, now and 
then in dyspepsia, in diseases of the lower extremities and 
hypochondria, in the convalescent stage of cholera, in 
Bright' s disease, but requires yet, in many cases, further 
confirmation. 

48. The urine in diabetes is usually very pale, of peculiar 
odor and high sp. grav., 1.030-1.052. Freshly passed, it 
very rarely gives a strong aeid reaction, usually neutral or 
feebly alkaline, but in consequence of fermentation rapidly 
becomes strongly acid in reaction, with the simultaneous 
formation of lactic, acetic, and traces of other volatile 
acids. 

Qualitative Detection of Sugar. 

49. Different methods serve for this purpose : — 

(1) The sugar can be obtained in a crystalline form, 
providing it occurs in considerable quantity in the urine. 



THE ABNORMAL CONSTITUENTS OF URINE. 55 

To this end, evaporate a portion of the urine to syrupy 
consistence, upon a water bath. The sugar separates from 
the solution upon standing, in yellow, warty masses, which 
by recrystallization can be further purified. Often there 
is found in urine a sugar which is perfectly uncrystallizable, 
and that remains in syrup form. 

(2) Moore's test. Place a quantity of urine in a narrow, 
tolerably long test tube, add sodium or potassium hydrate, 
and heat the upper portion of the liquid. If sugar be 
present in rather large quantity, this part will assume a 
yellow, or brownish red color, while the lower layer will 
retain its original color. 

(3) In doubtful cases the fermentation test is useful. A 
small portion of yeast is placed in a large test tube, and 
the latter then filled with the urine. The filled tube is in- 
verted over a small quantity of water, or urine, and allowed 
to stand for some hours, the temperature ranging from 
30-40°C. Any sugar present will break up into alcohol 
and carbon dioxide — 

C 6 H 12 6 = 2 C 2 H 5 HO + 2 C0 2 , 

Glucose 

and the resulting carbon dioxide will collect at the top of 
the tube. If the apparatus of Fresenius and Will be em- 
ployed in performing the test, the sugar can be estimated 
quantitatively. 

(4) Another test is to boil the urine under consideration 
for some time, with an ammoniacal silver nitrate solution. 
If there be any sugar present, the silver will deposit in 
metallic form, as a beautiful bright mirror upon the sides 
of the vessel. Formic and tartaric acids give a similar 
reaction. 

(5) A solution of indigo-carmine, rendered alkaline by 



58 CHEMICAL ANALYSIS OF THE URINE. 

75 parts dilute acetic acid (containing 30 per cent. acid). 
120 " water. 

(9) Trommer's test. Mix the sample of urine (freed of 
albumen) in a test tube, with a few drops of potassium or 
sodium hydrate, warm gently, to expel any ammonia 
present, filter if a large precipitate of earthy phosphates is 
formed, and then, after cooling, carefully add drop by drop, 
a dilute copper sulphate solution as long as the voluminous 
precipitate first formed dissolves. Heat the resulting clear 
blue liquid gently, and if sugar be present the solution will 
soon become cloudy, and instead of the blue color, yellow 
striped separations are noticed, which increase gradually 
until finally the entire liquid assumes a yellow color. On 
standing for a little time a yellow precipitate of hydrated 
cuprous oxide or of red cuprous oxide separates. Boiling 
should be avoided when heating the liquid. The heating 
of the urine with the alkaline hydrate before the addition 
of the copper solution should be very gentle, otherwise, 
when only traces of sugar are present, it can be so altered 
that it will not reduce the copper. If the alkaline copper 
solution and the urine are heated to boiling, the copper 
can be reduced by organic matters that are present and 
sugar be entirely absent. The inexperienced, therefore, 
should make repeated tests. 

If the preceding mixture of urine and alkaline copper 
solution is not heated at all, but left standing perfectly cold 
for 12-24 hours, if sugar be present, cuprous oxide will 
separate. (The other organic substances in urine only 
reduce the copper solution on the application of heat.) 

(10) Fehling's test. About 5 c.c. of Fehling's solution 
are poured into a test tube and brought to boiling. This 
should always be done before adding the suspected urine, 



THE ABNORMAL CONSTITUENTS OF URINE. 59 

for the reason that by standing for some time Fehling's 
solution undergoes decomposition, which unfits it for making 
the sugar test. If upon boiling a precipitate should form, 
the solution should not be used ; on the other hand, if no 
precipitate is formed, proof is shown that no change has 
taken place, and that the solution is reliable. The suspected 
urine is then added, drop by drop, and, if sugar be present, 
the blue color will change to green, and almost immedi- 
ately to yellow, hydrated cuprous oxide or red cuprous oxide 
being formed. If only minute quantities of sugar be present, 
several cubic centimetres of the urine may be required to • 
give the reaction. 

Blitz brings out sharply and elegantly the well known 
reaction between a solution of sugar and Fehling's liquid 
by mixing with the latter a concentrated sodium chloride 
solution, heating to boiling, and carefully adding to this a 
sample of the urine under examination. The strong sodium 
chloride solution prevents a mixture of the two liquids, so 
that at their point of contact the red coloration appears 
with great distinctness. 

Seegur (Centralblatt fur die Med. Wissenschaften, 1875, 
p. 323,) has confirmed the assertion that a solution rather 
rich in sugar will reduce Fehling's solution in the cold. 
This property is absent when the sugar is present in but 
minute quantities. He found' that an aqueous sugar solu- 
tion, containing 0.1 per cent, sugar produced scarcely any 
reduction in the cold ; that an aqueous sugar solution 
containing 0.05 per cent, sugar will not produce any re- 
duction whatever in the cold. An artificially prepared 
sugar solution containing 0.1 per cent, sugar caused in 
the cold a very slight decolorization of the copper solution 
without any separation of cuprous oxide. A sugar solution 



60 CHEMICAL ANALYSIS OF THE URINE. 

of the same strength (0.1 per cent.) after filtration through 
animal charcoal, was found entirely without action in the 
cold, while when warmed it caused the most beautiful 
separation of cuprous oxide. Experiments with pure uric 
acid solutions indicated that the same when containing as 
little as 0.5 per cent, uric acid reduced Fehling's solution 
very rapidly in the cold. 

Maly (Sitz. d. k. Akad. der Wissenschaft. Marz Heft, 
1871,) has found that 28 milligrams of a 1 per cent, crea- 
tinin solution dissolved the cuprous oxide furnished by 10 
milligrams sugar (1 per cent, solution). 

To detect sugar when contained in small quantities in 
urine, and also to free the latter from creatinin, Bence 
Jones employs the following modification* of Brucke's 
method : To 50 cubic centimetres of urine add 60 cubic 
centimetres of lead acetate solution (strength 10 percent.), 
filter, and to the filtrate add lead basic acetate as long as a 
precipitate forms, filter again, and to this last filtrate 
add ammonium hydrate. Collect the precipitate formed 
by the ammonium hydrate on a filter, wash thoroughly 
with water, remove with a horn spatula from filter paper 
and suspend it in water; through this mixture pass a 
stream of hydrogen sulphide. Filter off the precipitated 
lead sulphide, boil the filtrate, to expel the hydrogen 
sulphide remaining in solution, and after evaporating to a 
bulk equal to the original volume of urine employed or less, 
apply the tests for sugar. 

Another method proposed by Carnelutti and Valente 
(Gazz. Chim. x. 473-475) for the removal of creatinin is as 
follows: 100 c.c. of urine, decolorized by passing through 
animal charcoal, are evaporated to a syrup and mixed with 
1 c.c. of a solution composed of 25 per cent, zinc chloride, 



THE ABNORMAL CONSTITUENTS OF URINE. 61 

25 per cent, hydrochloric acid, 50 per cent, water. To the 
syrup, after the addition of the zinc chloride mixture, is 
added double the volume of alcohol, filtered, after standing 
several hours, the filter paper washed with alcohol, the 
alcoholic filtrate evaporated, and the residue diluted with 
water to the original volume of urine employed, and with 
this liquid the tests for sugar can be made. Fehling's 
quantitative method can be performed without any of the 
cuprous oxide going into solution. Loss of sugar does not 
take place in the performance of the above method. 

Small quantities of carbolic acid do not, but larger ones 
do affect the reaction of sugar with bismuth subnitrate. 
Carbolic acid also interferes in the test with Fehling's so- 
lution. Readily oxidizable substances, such as the hypo- 
phosphites, hinder the coloration of the sugar by potassium 
hydrate on warming (Moore's test), but hasten, apparently, 
the reduction of bismuth and copper. Hyposulphites also 
hasten the reaction with the bismuth, but deport themselves 
differently with Fehling's solution. On boiling the latter 
with hyposulphites, the blue color remains unaltered, and 
is decolorized on the addition of the sugar solution without 
separation of cuprous oxide. After standing awhile there 
is deposited a black mass, consisting mostly of copper sul- 
phide. Chloral, added to an alkaline solution of sugar and 
bismuth subnitrate, is rapidly decomposed, chloroform and 
formic acid are produced, and the reduction of the bismuth 
will be delayed until all the chloral is decomposed. 

Quantitative Determination of Sugar in Urine. 

50. (1) By fermentation. The carbonic acid apparatus of 
Fresehius and Will is employed here. It consists of two 
flasks connected by means of a glass tube bent twice at 



62 CHEMICAL ANALYSIS OF THE URINE. 

right angles. In one of the glass vessels we place about 30 
c.c. urine, together with some well washed yeast and a small 
quantity of tartaric acid . The apparatus is properly arranged , 
then weighed, and afterwards placed where there is a tem- 
perature of 20° to 30° C. In a short time fermentation 
sets in. The generated carbon dioxide passes through the 
sulphuric acid in the second flask and escapes into the air. 
In three days the fermentation is complete. The apparatus 
is then warmed gently and weighed when cool. The loss 
in weight, due to the escape of carbon dioxide, multiplied 
by 2.045 will represent the amount of sugar present in the 
given volume of urine. 

This method can be considerably modified, at least the 
apparatus can be dispensed with, by taking the specific 
gravity of a given volume of urine, adding a little yeast, 
allow it to ferment and again determine its specific gravity. 
Multiply the loss sustained by .23, or divide by 4.37, and 
the product will be the percentage amount of sugar present 
in the urine employed. 

(2) With the standardized alkaline copper solution. 
(The so-called Fehling's solution.) 

This is prepared as follows : — 

It is found that one molecule of sugar exactly reduces 
the copper in five molecules of copper sulphate, therefore, 
in order to make a copper solution in which we have suffi- 
cient copper sulphate to be exactly equivalent to five grams 
of sugar, we use the following proportion : — 

180 : J.247.5 ::5:x 

C 6 H 12 6 :5CuS0 4 +5H 2 :: 5grms.sugar : x= 34.6525 

grams crystallized copper sulphate, which are to be dis- 
solved in 200 c.c. water. 

173 grams chemically pure crystallized sodium potas- 



THE ABNORMAL CONSTITUENTS OF URINE. 63 

sium tartrate (Rochelle salts) are dissolved in 480 c.c. 
sodium hydrate solution of 1.14 specific gravity. To this 
we now gradually add, with constant stirring, the copper 
sulphate solution, and the mixed clear liquid is diluted with 
distilled water to one litre. Of this solution 
1000 c.c. = 5. grms. sugar. 
10 c.c. = .050 grm. sugar. 
10 c.c. of this copper solution will be reduced by 0.050 
grm. grape sugar. 

The above copper solution can only be preserved for a 
time without decomposition, by filling in small vessels of 
from one to two ounces capacity, which are then closed with 
tight fitting corks, sealed with wax or paraffin, and kept in a 
cool, dark cellar. Or the copper sulphate and double tar- 
trate solutions can be kept in separate, well corked bottles 
and mixed in proper proportion just before being used for 
analysis. However, it is always best to boil a sample of 
the Fehling's solution before using, to make certain that no 
decomposition has taken place, so that the copper will be 
reduced even in the absence of sugar. 

51. To make a sugar determination by this method a 
quantity of urine is diluted with nine or nineteen times its 
volume of water, and then placed in a burette. We now 
take 10 c.c. of the Fehling's solution, place it in a flask, or 
porcelain dish, dilute it with 40 c.c. water, and heat the 
mixture to boiling ; then allow the diluted urine to run in 
from the burette until all the copper has been reduced to 
cuprous oxide. This point is recognized when, after stand- 
ing some time, the cuprous oxide subsides, and the vessel 
held towards the light shows a colorless supernatant liquid. 
A filtered portion of this liquid acidified with acetic acid 
should not give a precipitate with ferrocyanide of potas- 



64 CHEMICAL ANALYSIS OF THE URINE. 

slum, nor with hydrogen sulphide. Another filtered por- 
tion is boiled with a few drops more of Fehling's solution. 
If a precipitate be formed in either of the first two tests, 
the reduction is not complete, and more urine must be 
added ; if the few drops of Fehling's solution added to the 
other portion be reduced, too much urine has been added, 
and the whole operation should be repeated. 

In making the test, it is advisable to heat the copper 
solution to gentle boiling, over a spirit lamp, or Bunsen 
burner, and when the solution assumes a red color, remove 
the flask, or dish, to allow the cuprous oxide to subside. 
The nearer the point of complete reduction, the more 
rapidly will the precipitate subside. As this determination 
is rather difficult for the. inexperienced, it should be re- 
peated several times. 

Albumen, as previously indicated, must be removed by 
coagulation and filtration. The calculation is as follows : — 

Suppose we diluted 10 c.c. of urine with 190 c.c. of water, 
and of this diluted liquid 25 c.c. were required to reduce 
the 10 c.c. of Fehling's solution, then we would have — 

200 : 10 : : 25 : x 

x = 10_X_25_2'50_ 19 * 00 
- 200" -200- V 5 C ' C - 

and in these 1.25 c.c. urine are contained 50 milligrams of 
sugar. From this we calculate how much sugar was elimi- 
nated in twenty-four hours. If a diabetic patient voided 
about 5000 c.c. urine, then we would have this proportion : 

1.25 c.c. : .050 milligram : : 5000 c.c. : x = 200,000 
milligrams, or 200 grams of sugar. 

(3) Knapp's Method yields results that agree perfectly 
with those obtained by the preceding method, and further, 



THE ABNORMAL CONSTITUENTS OF URINE. 65 

possesses decided advantages in the easy preparation and 
preservation of the mercuric cyanide solution employed. 
400 milligrams of the mercury salt require 100 milligrams 
of grape sugar for complete reduction ; 10 grams dry and 
pure mercuric cyanide are dissolved in enough water to 
effect solution, 100 c.c. of sodium hydrate solution of 1.145 
sp. grav. are added, and the whole diluted with water to 
one litre. In making an analysis, place 40 c.c. of the mer- 
curic cyanide solution in a flask, and heat to boiling. Now 
run in the urine so diluted as to contain about one-half per 
cent, sugar ; all the mercury is precipitated. In the quan- 
tity of the urine mixture required for the complete reduc- 
tion, there must have been exactly 100 milligrams of sugar. 

On adding the sugar solution to the boiling alkaline 
mercuric cyanide solution, the latter will become immedi- 
ately turbid, but clears again towards the end of the opera- 
tion and assumes a yellow color. To follow the course of 
the method, moisten a strip of Swedish filter paper, from 
time to time, with a drop of the mixture, and then with a 
glass rod bring a drop of ammonium sulphide close to the 
spot for about one-half minute. The whole spot at first 
becomes brown, but toward the end only its edge presents 
a clear brown ring, which may be noticed only by holding 
the transparent spot towards a bright light. Finally, the 
fresh, transparent spot is wholly unchanged by the ammo- 
nium sulphide, so that with some practice the T \ c.c. of a 
one-half per cent, sugar solution can be easily titrated. For 
complete satisfaction, filter finally a few c.c. of the liquid, 
acidify with acetic acid and test with hydrogen sulphide 
for mercury. 

(4) By polarization. The so-called observation tube is 
filled with clear, filtered urine, not containing any albumen, 



66 CHEMICAL ANALYSIS OF THE URINE. 

taking care, also, to prevent the inclosure of any air bub- 
bles, and then placed in Mitscherlich's or Ventzke-Soleil's 
polarization apparatus. Notice accurately on the scale 
and the verniers of the instrument, the rotatory power, and 
from this calculate the quantity of grape sugar by means 
of the formula — 

a 

in which p represents the quantity of sugar in grams for 
1 c.c. of urine; a, the observed rotation; 1, the length of 
the observation tube, and -f 56, the specific rotation. This 
method requires frequent practice, in order to obtain accu- 
rate results. 

Suppose we had, for example, found that the plane of 
polarization had been turned 3.5 to the right, then the 
equation would be — 

56 : 100 : : 35 : x 
100 X 35 



56 



= 6.25 



therefore, a rotation of 3.5 degrees would indicate 6.25 per 
cent, sugar. 

INOSITE IN URINE. 

52. Inosite has been found constantly in urine in B right's 
disease and albuminuria, in uraemia after the use of dras- 
tics, in diabetes mellitus, in two cases of carcinoma, and 
once in the urine of a convalescent from cholera. In one 
instance of diabetes the inosite gradually displaced the 
sugar originally present. Kiilz (Centrallblatt f. d. med. 
Wissensch., 1876, p. 550.) has confirmed the assertion of 
Strauss according to whom inosite is a constituent of nor- 
mal urine, whenever there has been excessive drinking of 



THE ABNORMAL CONSTITUENTS OF URINE. 67 

water. It may be detected as follows : Urine from which 
albumen has been completely removed is saturated with 
lead acetate solution, filtered, and the concentrated filtrate 
mixed with basic lead acetate as long as a precipitate appears. 
The latter contains the inosite combined with it. The pre- 
cipitate is collected on a filter paper and well washed with 
water, and then scraped off and suspended in water, and a 
stream of hydrogen sulphide passed through. The pre- 
cipitated lead sulphide is filtered off. The filtrate from the 
lead sulphide may deposit some uric acid. This can be fil- 
tered off, the filtrate concentrated quite considerably, and 
while boiling mixed with three to four times its volume of 
alcohol. Should this produce a heavy precipitate which 
tends to adhere to the sides of the vessel, then pour off the 
alcoholic solution ; but if there is only a flaky turbidity, 
filter through a warmed funnel, and allow the solution to 
cool. In about twenty-four hours the inosite will separate 
out from the filtrate in cauliflower-like grouped crystals. 

Inosite is insoluble in alcohol and ether, readily soluble 
in water. The aqueous solution has a sweet taste. Yeast 
does not decompose inosite into alcohol, but decaying 
cheese will effect this. It is further recognized in its be- 
havior toward nitric acid. On evaporating it with nitric 
acid to dryness, and moistening the residue with a little 
ammonium hydrate and calcium chloride, and again evap- 
orating, a brilliant rose-red coloration results. A trans- 
parent gelatinous mass, which soon becomes starch-like in 
appearance, is produced on warming an inosite solution 
with basic lead acetate. The reaction with mercuric nitrate 
is also worthy of note. 



68 CHEMICAL ANALYSIS OF THE URINE. 

LACTIC ACID AND LACTATES. 

53. Lactic acid has been observed in urine in the acid fer- 
mentation, and results, very likely, from the decomposition 
of urinary extractive and coloring matters. It is also as- 
serted that this acid has been found in urine when there 
was obstruction of the oxidation in the blood, therefore, in 
disturbances of respiration, digestion and nourishment in 
the urine of rachitic children and in leucaemia. 

As lactic acid does not present any marked chemical 
properties, its zinc salt, which crystallizes readily in char- 
acteristic forms — mallet-shaped — is used to detect it. The 
urine intended for its preparation should be as fresh as 
possible. Inasmuch as its occurrence in urine is very 
variable, and it does not afford any definite diagnosis, we 
can omit the remaining properties. 

FATS AND FATTY ACIDS. 

54. Fat is very rarely found in urine. It has been 
noticed in the fatty degeneration of the kidneys (Bright's 
disease), in the fatty degeneration of the epithelial cells of 
the urinary organs and bladder, and in excessive chylous, 
or fatty blood serum (urina chylosa, cause unknown). 
From time to time, of the volatile acids, butyric has been 
found, and in combination in fermented diabetic urine, 
acetic and propylic acids. Owing to the small quantities 
of fat in urine, it is very difficult to detect by chemical 
means. The microscope affords us the best solution of the 
problem, as the fat globules appear here as flattened round 
plates of remarkable refracting power, and dark, tolerably 
irregular contours. When it is impossible to recognize the 
fat under the microscope, the urine under examination is 



THE ABNORMAL CONSTITUENTS OF URINE. 



69 



evaporated upon a water bath, the residue dried for some 
time at 110° C, then extracted repeatedly with ether. 
When the ether has evaporated, only fat remains, and its 
presence can now be confirmed under the microscope, and 
by its deportment toward heat (acrolein) and paper (grease 
spots). 

BILIARY COLORING MATTERS, BILIARY SALTS AND TAURIN. 

55. Although biliary coloring matters are likely to occur 
in healthy persons during the hot portions of the year, such 
instances are rare. Both biliary coloring matters and 
salts, and now and then taurin (decomposition of tauro- 
cholic acid), are found in icterus. Urine charged with 
biliary pigments is easily recognized by its decided tinge 
of color, being at one period red-brown, and then grass 
green. Such urine foams strongly when shaken, and colors 
filter-paper yellow or green. 

For the detection of either of the above we employ nitric 
acid. Place in a test tube some concentrated and slightly 
yellow-colored nitric acid, and then carefully add, by 
means of a pipette, some of the urine under examination, 
taking care that the two liquids do not intimately mix. In 
the presence of biliary pigments there will be produced at 
the junction of the two liquids a beautiful play of colors, 
at first a beautiful green ring, which gradually rises higher, 
exhibiting slowly at its lower surface a blue, violet, red and 
finally yellow ring (green is characteristic for bile pig- 
ments). 

Urine containing bile, when treated with hydrogen per- 
oxide, ferric chloride, and an acetic or phosphoric acid 
solution of lead superoxide, shows a beautiful green color. 

Masset (Journ. de Pharm., et de Chim., [4], 30, 49), 



70 CHEMICAL ANALYSIS OF THE URINE. 

employs the following modification of Gmelin's test for the 
detection of biliary pigment in urine. 2 cubic centimetres 
of urine are acidified with 2-3 drops concentrated sulphuric 
acid, and then a small crystal of sodium nitrite introduced 
into the liquid. In the presence of bile pigments magnifi- 
cent grass-green streaks appear, which, on shaking, color 
the entire liquid dark green. This color does not disappear 
on boiling and remains many days unaltered. Even traces 
of biliary coloring matter produce a distinct pale green 
coloration. 

Traces of bilirubin are detected by shaking the urine 
with chloroform, which becomes yellow in color If nitric 
acid (containing nitrous acid) be poured on the chloroform 
the play of colors mentioned before as produced with nitric 
acid will be noticed. 

To detect the acids of the bile (of which cholic acid is 
the starting point), separate the sodium salts from the 
urine and treat the concentrated aqueous solution with 
from 2 to 3 drops of sugar solution (1 to 4) then add a 
little pure concentrated sulphuric acid. The liquid is at 
first turbid, then it becomes clear and almost at the same 
moment yellow, then pale cherry red, dark carmine red, and 
finally beautiful purple violet. 

LEUCIN, TYROSIN AND CYSTIN. 

56. Leucin and tyrosin have been found in acute yellow 
atrophy of the liver, in typhus, variola, and in the urine of 
an epileptic after injury to the spinal cord. Urine con- 
taining cystin has frequently been observed. The relation 
of cystin to any definite changes in disease has not yet been 
determined. When leucin and tyrosin are abundant in 
urine, they can easily- be detected. Tyrosin is either al- 



THE ABNORMAL CONSTITUENTS OF URINE. 71 

ready found crystallized out, or it separates simultaneously 
with leucin on evaporating the urine to a small bulk and 
allowing it to cool, when the well known characteristic 
forms are microscopically recognized (leucin in brown, oily- 
like layers, tyrosin in sheaf-like needles). If the quantity of 
these substances is not so abundant that they appear upon 
the evaporation of the urine, the method of Frerichs 
should be pursued. A rather large quantity of urine, 
usually rich in biliary pigments and albumen, is precipi- 
tated with basic acetate of lead, filtered, the excess of lead 
in the filtrate removed by passing a stream of hydrogen 
sulphide through it, and the filtered and clear solution 
reduced to a small volume on a water bath. If tyrosin 
is present, in twenty-four hours it will be found nicely 
crystallized out ; while the leucin, being much more 
soluble, separates later. 

OCCURRENCE OF FIBRIN IN URINE. 

57. Fibrin very rarely occurs in urine, and is of little 
definite diagnostic importance. When it is found present 
we are justified in the conclusion that there has been a 
fibrinous transudation from the blood into the kidneys or 
urinary passages. The presence of fibrin is characterized 
by the formation of fibrinous coagula some hours after the 
urine has been voided. These coagula deposit as a 
sediment or convert all the urine into a gelatinous mass. 
The microscope will show the regular fibrin cylinder as 
rolled-up with sharp contour and yellow or brown yellow 
color. (Plate hi, Fig. 2.) 

BLOOD PIGMENTS IN URINE. 

58. Blood pigments have been detected in urine in cer- 



72 CHEMICAL ANALYSIS OF THE URINE. 

tain diseases which accompany dyscrasia and blood de- 
generation, in scurvy, in putrid typhus fevers, in per- 
nicious alternating fevers, and after the inhalation of 
hydrogen arsenide. 

In these instances the urine is bloody, colored from red- 
brown to ink black. Yet a microscopic examination will 
not reveal the elemental forms of the blood. 

Upon boiling such urine alone, or after the careful addi- 
tion of some drops of acetic acid, a brown-red coagulum is 
formed, which, with alcohol containing sulphuric acid, 
yields hsematin. 

BLOOD IN URINE. 

59. In troubles induced by the presence of calculi in the 
bladder or kidneys, causing a mechanical lesion of certain 
vessels, or in violent desquamative nephritis, finally, in 
severe cystitis in which the texture of the bladder suffers, 
blood can occur as such in the urine. It can, in addition, 
occur as a result of the effusion of the blood into the uri- 
nary canal, that, by the coagulation of the blood of the 
urethra the passage for the urine will be obstructed so 
that the voiding of the urine will be impaired, or that these 
coagula will induce the formation of permanent concre- 
tions in the urinary channel. If blood be present in the 
urine, fibrin and albumen, as integral parts of the blood, 
will also be found, and, therefore, it will be very necessary 
to proceed carefully if we wish to ascertain whether all the 
albumen occurring in the urine originated from the effused 
blood or from other sources. 

Almen (Neues Jahrbuch fur Pharmacie, 40 p. 232,) 
recommends the following for the detection of blood in 
urine. Mix in a test tube some drops of tincture of guaia- 
cum with an equal volume of oil of turpentine, and shake 



THE ABNORMAL CONSTITUENTS OF URINE. 73 

until an emulsion forms, then carefully add the urine under 
examination, so that it falls to the bottom of the tube. On 
agitating the emulsion with the urine, the guaiacum resin 
is rapidly precipitated as a white, afterward dirty yellow 
or green precipitate. If there be blood in the urine, and 
even if only in traces, the resin is colored a more or less 
intense blue, often almost indigo blue in color. In normal, 
albuminous, or urine containing pus, this blue coloration 
does not occur, but only appears in the presence of blood. 

HYDROGEN SULPHIDE IN URINE. 

60. Hydrogen sulphide is very rarely observed in urine. 
It has been noticed in the so-called reabsorbed urine, and 
its occurrence attributed to the fusion of the exuded 
proteids. According to Beetz, under certain conditions 
ammonium sulphide can reach the blood from the skin, 
and there produce phenomena of poisoning similar to those 
observed in the inhalation of sewer gas. In this case the 
urine yields tests for ammonia and hydrogen sulphide. In 
violent cystitis from the decay of albuminous urine in the 
bladder, hydrogen sulphide will be formed, and it is then a 
very unfavorable prognosis. The detection of it is easy — 
the odor distinguishing it. A slip of paper moistened with 
lead acetate is immediately blackened when immersed or 
held over urine containing hydrogen sulphide. In this 
test the urine should be slightly warmed. 

OXYMANDELIC ACID. 

61.0. Schultzen and L. Riess discovered oxymandelic acid 
as an abnormal constituent in urine, together with leucin, 
tyrosin, and sarco-lactic acid. Its formula is C 8 H 8 4 . 

F 



74 CHEMICAL ANALYSIS OF THE URINE. 

The urine in which this occurs contains also biliary pig- 
ments, biliary acids, albumen in traces, and that peptone- 
like substance which is noticed in urine in considerable 
quantities after phosphorus poisoning. The urea is either 
perfectly absent, or present in diminished quantity. 

To obtain the acid, free the urine, by evaporation, from 
tyrosin and leucin, precipitate the mother liquor with 
alcohol, evaporate the alcoholic solution, and the syrupy 
residue, after the addition of dilute sulphuric acid, is ex- 
hausted completely with ether. The united ethereal ex- 
tracts leave upon evaporation a brown, liquid residue, from 
which long, thin, colorless needles separate, which are 
then dissolved in water and the solution filtered. In the 
feebly yellow-colored nitrate lead acetate produces only a 
slight nocculent precipitate, which decolorizes the liquid. 
With basic lead acetate, the filtrate gives a voluminous 
nocculent precipitate, which condenses to a heavy, granu- 
lar, crystalline powder. This compound is suspended in 
water and decomposed by hydrogen sulphide. On evapo- 
ration the nitrate deposits colorless, silky, very flexible 
needles — constituting the new acid. 

INDICAN. 

62. Indican has recently been found to be an indoxyl- 
sulpho acid. Jaffe estimates it quantitatively by means 
of bleaching lime. 1000 to 1500 c.c. of urine are made 
alkaline with calcium hydrate and the phosphates then re- 
moved by means of calcium chloride. Filter after twelve 
hours, evaporate the filtrate to a thick syrup. The syrupy 
residue is warmed some minutes with about 500 c.c. alcohol, 
then brought into a beaker and allowed to stand twelve or 
twenty-four hours. Filter and distill off the alcohol. The 



URINARY DEPOSITS. 75 

residue is dissolved in a large quantity of water and preci- 
pitated with a very dilute solution of ferric chloride. The 
nitrate from the iron precipitate is mixed with ammonium 
hydrate, boiled, and after filtration evaporated to 200- 
250 c.c. With this solution the determination is made. 
First determine the amount of chloride of lime necessary 
to separate the indigo. To this end measure out 20-40 c.c. 
of the liquid, and dilute this gradually with definite amounts 
of water, until 10 c.c. of the mixture treated with an equal 
volume of hydrochloric acid show a perceptibly blue color- 
ation on the addition of a drop of a saturated bleaching 
lime solution. Multiplied experiments have shown that 
the number of volumes of dilution which can be added to 
an indican solution until the appearance of the limit of the 
reaction, is about double the number of drops of the bleach- 
ing lime solution, which will show the maximum indigo 
yield for 10 c.c. of indican. When the right proportion 
has been determined, mix 200-300 c.c. of urine with bleach- 
ing lime and hydrochloric acid, allow to stand at least 
twelve hours, collect upon a filter that has been extracted 
with hydrochloric acid and washed, dried, and weighed. 
Dissolve out the hippuric and benzoic acids with water, 
wash the residual indigo with dilute ammonium hydrate, 
and finally with water, dry, precipitate and filter at 105 to 
110° C, and weigh. 

V. 

URINARY DEPOSITS (SEDIMENTS). 

63. Urinary deposits are solid, undissolved substances 
in the urine which at first are mostly suspended in the 
latter, but after shorter or longer periods form a precipi- 
tate. Some are produced after, others before, the urine has 



76 CHEMICAL ANALYSIS OF THE URINE. 

been voided. In the latter case they may form in the 
tracts of the urine (urinary organs, bladder, etc.), and 
under favorable circumstances produce calculi. 

Many urinary sediments whose constituents were at first 
dissolved, separate or form in consequence of the peculiar 
alterations of the urine. These have already been de- 
scribed in preceding paragraphs, under the name of acid 
and alkaline urine fermentation. 

64. The microscope is an indispensable aid in the exami- 
nation of these deposits. Without it we would, in many 
instances, not be capable of arriving at a correct, conclusive 
decision. By its assistance we distinguish the various sedi- 
mentary forms, classing them as amorphous, crystallized 
and organized bodies. These are, however, not alike for 
every reaction of the urine. So far as the organized bodies 
are concerned it is only partially correct, while the occur- 
rence of the crystalline and amorphous bodies is dependent 
in part upon the reaction. 

65. Depending on the reaction in urine we find various 
substances in the deposits: — 

A. In acid urine, are present : — 

(a) amorphous bodies : urates, phosphates and fats. 

(b) crystalline bodies : calcium oxalate, uric acid, cal- 
cium phosphate, cystin, tyrosin, hippuric acid. 

(c) organized bodies : mucous coagula, mucous corpus- 
cles, pus, blood corpuscles, urinary casts, epithelial 
cells, fermentation and thread fungi, vibrionse, sper- 
matozoids, cancerous tissues, sarcina ventriculi 
Goodsir. 

B. In alkaline urine are found : — 

(a) amorphous bodies: calcium carbonate, calcium 
phosphate. 



URINARY DEPOSITS. 77 

(b) crystalline bodies: magnesium ammonium phos- 
phate (triple phosphate), ammonium urate, crystal- 
lized calcium phosphate. 

(c) organized bodies : in addition to the above, infu- 
sorise and confervse (fermentation and thread-like 
fungi are increased). 

Therefore, before beginning a microscopic examination, 
observe whether the urine has been newly voided, the re- 
action whether alkaline or acid. This done, the sediment 
is allowed to subside, the supernatant liquid decanted, and 
by means of a pipette, a drop of the sediment is placed on 
a glass slide, covered with a glass circle, and then brought 
under the objective of a microscope. Move the slide about, 
until all points have passed the field of vision. Having 
examined one sample, take a second, and be it noticed 
here that this specimen be taken from different layers of 
the sediment, inasmuch as some substances deposit more 
rapidly than others, and many, like calcium oxalate, only 
after the expiration of several hours. If filtration had 
been necessary for the separation of the deposit, be careful, 
in cleaning the filter paper, not to bring any fibres of the 
paper under the microscope and consider them solid con- 
stituents of the sediment. 
I. The urine reacts acid. 

A. The entire sediment is amorphous, presenting par- 
tially irregular masses, partially moss-like intertwined 
series, consisting of extremely fine grains. 

Carefully warm the drop on the object glass, 
(a) Perfect solution follows = urates. As a confirma- 
tory test, add, after cooling, a drop of hydrochloric 
acid and allow to stand from one-quarter to one- 
half hour. If rhombic tablets of uric acid form in 



78 CHEMICAL ANALYSIS OF THE URINE. 

this time, proof is sufficient. Usually this sediment 
consists of acid sodium urate, and is distinguished 
by a more or less red color. 

(b) The sediment does not dissolve on the application 
of heat, but dissolves in acetic acid without efferves- 
cence = calcium phosphate. Chemically, the calcium 
is proven by ammonium oxalate ; the phosphoric 
acid by ammonium hydrate, and magnesium sul- 
phate forming ammonium magnesium phosphate, 
or by means of ammonium molybdate. 

(c) Beneath the sediment are found drops which re- 
fract light strongly = fat. 

B. The sediment, or deposit, co?itains well formed crystals : — 

(a) Calcium oxalate. Minute, shining, perfectly trans- 
parent, quadratic octahedra in the form of envel- 
opes, which refract light strongly (Plate n, Fig. 2). 
As these crystals are light in weight, they deposit 
very slowly, and can readily be overlooked by the 
inexperienced. The urine should be allowed 12-24 
hours to deposit and then be carefully decanted. 

(b) Uric acid, following its principal form, crystallizes 
in rhombic tablets with rounded, blunt corners, 
which are known as Wetzstein's form (Plate i, Fig. 
5). The crystals may be very small and some very 
complicated, building themselves upon accidental 
impurities, e. g., threads, and forming series of hairs 
and long cylinders. 

Again, the crystals are greatly developed, and 
united to a nucleus, when they appear upon the 
edge (fan-like), or upon the plane (shingle-like). 
Uric acid has also been found in cask shapes and 
long spears, combined with rosettes. Owing to 



URINARY DEPOSITS. <» 

coloring matters precipitated at the same time, the 
uric acid is either pale yellow, or brown-red to dark- 
brown (Plate i, Figs. 4-5). 

For chemical confirmation, see page, 32. 

(c) The crystallized calcium phosphate, viewed under 
the microscope, presents either individual keel- 
shaped crystals, or several are arranged in regular 
order, so that they are with their sides towards each 
other, and their ends converging to one point. In 
addition, perfect circular rosettes are found, and 
sometimes the crystals not only arrange themselves 
in circles, but build parts of spheres. The urine 
usually reacts feebly acid. 

(d) Cystin forms regular, six-sided tables, soluble in 
ammonium hydrate and hydrochloric acid. They 
carbonize and burn on being heated. Boiled with 
a sodium hydrate solution of lead oxide, lead sulph- 
ide is produced. The chemical test for cystin con- 
sists in this last experiment, and that on being 
heated on platinum foil it does not fuse, but burns 
with a greenish blue flame and the diffusion of an 
odor very similar to prussic acid. 

Cystic urine is generally pale. The assertion has 
been made that, frequently, several individuals of 
the same family will suffer from cystinuria. 

(e) Tyrosin forms delicate short needles, which cross 
each other so frequently that they present a sheaf- 
like appearance, of which every two sheaves super- 
impose in the form of a cross. Chemically the 
tyrosin crystals are tested according to the method 
of Piria, or Hoffmann. According to the first, a 
minute quantity of the sediment is placed on a watch 



80 CHEMICAL ANALYSIS OF THE URINE. 

glass and moistened with two to three drops of sul- 
phuric acid. In about half an hour, add a little 
water, neutralize with sodium carbonate, as long as 
it effervesces, then filter. If the sediment was ty- 
rosin, the solution, on the addition of neutral ferric 
chloride, will show a violet color. Hoffmann's 
method is simpler. Pour water over a portion of 
the sediment, boil, and add to the boiling liquid a 
few drops of mercuric nitrate, when a red precipi- 
tate will form, while the supernatant liquid is colored 
rose to purple red. 

Urine containing tyrosin frequently contains bili- 
ary pigments. 
(f) Hippuric acid, as a sediment, rarely occurs. It 
crystallizes in needles and rhombic prisms, soluble 
in water. (See Plate i, Fig. 3.) 
C. The sediment contains organized bodies: — 

(a) Mucous coagula, forming wound-up strips, consist- 
ing of serrated, very minutely arranged points and 
grains, frequently accompanied by sodium urate. 

(b) Mucous Corpuscles. Very small, contracted and 
granulated corpuscles, generally combined at the 
edges to large shield-like groups. 

Large quantities of mucus can form in urine, 
without affecting the transparency of the latter. 
Only on protracted standing, when there commences 
a deposition of urates, or when the urine contains 
more epithelium than usual mixed with it, does the 
mucus become visible, as a cloud. If the turbidity 
disappears on the application of heat, the urates were 
the cause. Small crystals of calcium oxalate and 
uric acid, as well as individual mucous corpuscles, 



URINARY DEPOSITS. 81 

or epithelium of the bladder, which bodies had been 
suspended in the mucus, have also been found. 

(c) Blood corpuscles form circular, slightly bi- con cave 
disks, generally with a yellow appearance, again 
reddish with a faint touch of green. They greatly 
expand by acetic acid, dissolving in this more or 
less slowly. (See Plate in, Fig. 3.) 

Particular attention should be directed to swollen, 
spherical and also distorted zigzag forms (readily 
produced by a concentrated sodium sulphate solu- 
tion). 

In the presence of blood the urine contains al- 
bumen. To detect blood pigments in urine, pre- 
cipitate the earthy phosphates of the urine in a test 
tube with potassium hydrate, warming gently. In 
the precipitation the phosphates carry down the 
pigments, appearing not white, as in normal urine, 
but blood red. When but a small amount of blood 
pigment is present in the urine, the earthy phos- 
phates show dichroism. 

(d) Pus. Round, pale, granulated cells of varying 
magnitude, usually as large again as blood corpus- 
cles, increasing markedly when touched with acetic 
acid, and losing their granulated surface and giving 
rise to residues of varied forms and groupings. It 
is impossible, either chemically or microscopically, 
to distinguish these corpuscles from mucous corpus- 
cles, but in the presence of pus the urine always 
contains albumen. (See Plate in, Fig. 4.) 

By Donne's test the pus in urine can be detected 
without the assistance of a microscope. To do this, 
pour off the urine from the sediment, add a small 



82 CHEMICAL ANALYSIS OF THE URINE. 

piece of solid potassium hydrate to the latter, and 
stir some minutes with a glass rod. If the sediment 
consists of pus, it will be deprived of its white color, 
becoming greenish and glassy, at first thready, 
finally more compact, until eventually it results in 
a coherent body, i. e., it has assumed the appearance 
peculiar to pus in strong ammoniacal urine. Only 
in case the quantity of pus was small, it cannot be 
expected to result in a compact lump, but the sedi- 
ment may be made to disappear, and a thready, 
gluey liquid results. 

(e) Urinary casts are tube-like cylinders, often accom- 
panied by blood and pus corpuscles, holding in their 
substance or walls epithelial cells and mucous cor- 
puscles. 

(a) The epithelial casts of the Bellini tubes, whose 
round cells are distinctly visible as a delicate 
molecular mass. 

(/5) Granulated kidney casts are of granular, 
cloudy appearance. 

(j) Hyaline kidney casts are solid, of paler, more 
transparent appearance. Often distinguished 
from the surrounding liquid with only the greatest 
difficulty. (See Plate in, Fig. 1.) 

(f ) Epithelial cells in their different forms, dependent 
on their origin. (See Plate n, Fig. 6.) 

(1) Squamous epithelium. Round, longitudinal or 
polygonal cells from the major and minor labise 
and the vagina, from the female urethra, the 
bladder, the kidneys. (See Plate in, Fig. 6.) 

(2) Cylindrical and spheroidal epithelium from the 
lower layer of the mucous membrane of the bladder. 



Plate II 



Fig. 1. 




Uric Acid, Sodium Urate and fermentation funj 

Fig. 3. 




Ammonium Urate. 

Fig. 5. 





Ualcium Oxalate. 

Fig. 4. 




Triple Phosphates 



Epithelial casts and Epithelial cells. 



URINARY DEPOSITS. 83 

(3) Glistening columnar epithelium from the uterus. 
(Addition of iodine solution makes all these for- 
mations more distinct under the microscope), 
(g) Fermentation and Thread-like Fungi. In the first 
stage of the acid urine fermentation they accompa- 
ny the sediments of sodium urate, free uric acid and 
calcium oxalate, but are found most frequently in 
diabetic urine, and such as has passed into ferment- 
ation. 

(1) The fermentation fungi form small nucleated 
cells, which increase by formation of sprouts, and 
thus form simple or intertwined series. 

(2) Thread-like fungi often produce so thick a 
tissue that they obscure the field of vision. 

(h) Vibrionse are short, delicate rods, moving actively 
hither and thither (under high power observed in 
feebly acid and alkaline urine). 

(i) Spermatozoids. Microscopic, somewhat elongated, 
pear-shaped bodies, with a more or less long, hair- 
like tail, which may or may not be in constant vi- 
bration. They are found — 

(1) After coition. When a portion of the seminal 
fluid had remained in the urethra and was dis- 
charged in the urine later. 

(2) In spermatorrhoea. Besides the independent 
disease of this name, involuntary emissions of semi- 
nal fluid have been noticed in serious cases of 
typhus. 

(j) Cancerous masses: 

(1) Distinct cancer cells. 

(2) Small pieces of cancerous tissues. 

The first are often unusually large, most 



84 CHEMICAL ANALYSIS OF 1 HE URINE. 

frequently having, apparently, a cilium with very 
large, often multiplied, nuclei. Care must be 
taken not to confound the ciliated cells originating in 
the pelvis of the kidney with the cancerous cells. 
The superstructure of the villous cancer consists of 
dendritic vegetation, upon which sometimes the 
epithelial growth rests. Such masses are voided 
spontaneously with the urine from the bladder. 
Again, it is only after examination, as, for instance, 
in the introduction of the catheter, that they are 
loosened and appear subsequently in the urine. 
(See Plate in, Fig. 5.) 
(k) Sarcina ventriculi Goodsir. Very rare. The 
characteristic form is not readily confounded with 
anything else. 
II. The urine reacts alkaline. 

A. The sediment contains amorphous bodies. 

(a) In alkaline urine these consist only of calcium 
phosphate. 

B. The sediment contains crystals. 

(a) The ammonium magnesium phosphate occurs 
usually in combinations of the rhombic, vertical 
prisms, in equal coffin-lid-like crystals, which acetic 
acid dissolves easily (distinction from calcium oxa- 
late), and on warming with sodium hydrate, am- 
monia gas is liberated. (See Plate n, Fig. 5.) 

(b) Ammonium urate consists of brown colored spheres, 
which are developed singly, or every two are com- 
bined to double spheres, presenting entire conglo- 
merations with reniform surface. The latter is 
smooth or set with small points like a thorn apple, 
or the projecting points are long, evenly divided, 



URINARY DEPOSITS. 85 

and then mostly bent, which gives rise to a great 
multiplicity of intermingled forms. (See Plate n, 
Fig. 3.) 

Ammonium urate, like other urates, gives the 
murexide test. 
C. The sediment contains organized bodies : 
Besides blood, mucus and pus corpuscles, fermenta- 
tion and thread-like fungi, infusorise and confervas 
are found. 

Relations of Sediments to the Diagnosis of Disease. 

66. (1) Uric acid and urates occur not only in pathological 
urine, in acute, febrile diseases, but also in normal 
urine. In newly voided urine sediments of free uric 
acid never occur, except in renal calculus, while on 
the other hand every urine in the course of acid fer- 
mentation deposits uric acid crystals. The deposits 
of urates, especially potassium and sodium urates, are 
very frequent, and represent the fever sediments (sedi- 
menta lateritia), long known to physicians. They are 
sometimes deceptively similar to mucus, pus and blood, 
and are only recognized by their microscopic char- 
acter. 

(2) Deposits of calcium oxalate occur in both healthy 
and diseased individuals. Oxaluria, which is the 
name applied when they occur abundantly, is of great 
diagnostic importance, although it occurs in some 
other diseases, as dyspepsia, spermatorrhoea and dis- 
eases of the spinal cord. In oxaluria the urine is dark 
in color. (See Plate n, Fig 2.) 

(3) Hippuric acid deposits are found frequently after 
the eating of fruit, the ingestion of benzoic and cinna- 



86 CHEMICAL ANALYSIS OF THE URINE. 

mic acids, and in various diseases. It has very little 
diagnostic importance. 

(4) The rarely observed sediments of cystin are of little 
diagnostic value. Generally present in renal calculus. 

(5) Tyrosin sediments have been observed in acute liver 
diseases. 

(6) Sediments of ammonium magnesium phosphate 
(triple phosphate) are found constantly, when the urine, 
because of the conversion of urea into carbon dioxide 
and ammonia, becomes alkaline. 

(7) Calcium phosphate occurs under the same condi- 
tions. 

(8) Mucus corpuscles (mucin) are constantly present in 
traces in normal urine, also in febrile conditions of the 
most varied type, as pneumonia, pleuritis, typhus, res- 
piratory and intestinal catarrh, meningitis, etc. 

(9) Tube casts are observed in many diseases, particu- 
larly Bright's disease of the kidneys. They constitute 
the principal basis in the diagnosis and prognosis of 
certain diseases of the renal parenchyma. 

(10) Spermatozoids exist in urine after pollution or 
coitus ; also not unfrequently in the urine of typhoid 
patients. They point to an unusual and decidedly 
excessive irritation of the genital organs. 

(11) Fungi and infusorise in freshly voided urine in- 
dicate that it has decomposed in the bladder, which is 
tolerably often the case in catarrh of the bladder. 

(12) Pus in urine always indicates suppuration in the 
uropoetic system, or points to an abscess related to the 
latter. The question of importance is, is the pus the 
product of a superficial affection of the mucous mem- 
brane (catarrhal inflammation), or of a graver affec- 



EXAMINATION OF URINE. 



87 



tion of this membrane, intimately connected with 
material alterations ? To answer the question observe 
the continuance of the suppuration, and the properties 
of the pus. 
(13) Cancer and tubercular masses show the presence 
of cancerous or tubercular depositions which have 
softened in almost any part of the uropoetic system : 
example, cancer of the bladder, and rarely, cancer of 
the kidneys. 

VI. 

PRACTICAL HINTS TO A COURSE FOR THE QUALITATIVE 
AND QUANTITATIVE EXAMINATION OF URINE. 

67. As a rule, it is scarcely necessary to examine for all 
the normal and abnormal substances in urine. Proof of 
the presence of one or several of the mentioned constitu- 
ents is sufficient for diagnosis, and it is only where the 
physician desires an accurate knowledge of all the nourish- 
ment relations of an individual, that it can be of value 
to him to extend the analysis to all substances found in the 
urine. In such instances, a single analysis is insufficient, 
only a series of repeated analyses being satisfactory. 

The substances that are to be looked for dictate the 
course of analysis. 

In examining urine, we always regard it as pathological, 
consequently, we search for abnormal constituents. An 
exception would be urine containing a sediment. Here 
examine both the liquid and the deposit, and class the sub- 
stances found as first (a) in the sediment, (6) in solution. 

Of course, some of the normal constituents should be 
searched for, such as salts, etc., and in the report of the 



00 CHEMICAL ANALYSIS OF THE URINE. 

examination, catalogue the various ingredients found under 
the headings normal constituents, and abnormal constitu- 
ents. This is advisable for the practitioner, because he 
does not always retain in memory the various constituents 
of urine, so that from an arbitrary arrangement of the 
normal and abnormal constituents, he is able to present a 
clear picture. This is more readily accomplished when the 
detected substances are arranged under the mentioned 
headings. This is advantageous, too, where an accurate ex- 
amination may be required. 

The physician having determined the substance to look 
for, whose chemical detection is principally concerned, the 
same is sought under the respective headings, and tested as 
therein directed. 

If, on the contrary, a general examination is desirable, 
pursue the plan recommended by Neubauer. 

68. Qualitative course. 

I. Determine the reaction with litmus. 

The urine may be : — 

(a) acid and clear. 

(b) acid and sedimentary. 

(c) neutral or alkaline. In the latter case, a deposit 
is usually present. The filtered urine, free from 
sediment, is further tested. (Section 65.) 

II. Albumen. Biliary pigments and blood. Heat a 
small quantity of the urine (if it does not give an 
acid reaction); with addition of a drop or two oi 
acetic acid, to boiling. The formation of a coagulum, 
not removed by nitric acid, indicates albumen. If 
the coagulum is — 

(a) white: it consists of pure albumen. (See page 52.) 

(b) greenish: there is good reason to suspect biliary 



Plate TIL 



Fig. 1. 




Fig. 2. 



Blood corpuscles. 

Fig. 5. 





Fine granulated casts. 

Fig. 4. 




Organized growth found in urinary sediment from 
ail individual having cancer in the bladder. 



Sediment from normal urine, showing several 

mucus corpuscles (young cells) and 

squamous epithelia. 



EXAMINATION OF URINE. 89 

pigments, especially if the urine be highly colored. 
(See page 71.) 
(c) brownish-red: blood may be present. (See page 72.) 

III. Urea. Creatinin. Uric acid. Hippuric acid. 
Lactic acid. Earthy phosphates, etc. About 400 to 
500 c.c. of clear urine, free from sediment and albu- 
men coagulum, are evaporated upon a water bath to 
thick syrupy consistence, and then divided into two 
parts (i and f ). 

(1) i of the residue is exhausted with strong alcohol ; 
allow the undissolved portion to subside, filter, wash , 
the residue again with strong alcohol and test the 
solution according to a and b, the residue according 
to 3. 

(a) Urea. A small portion of the alcoholic liquid is 
evaporated almost to dryness on a water bath, the 
residue is dissolved in a little water, and a few drops 
of pure nitric acid free from nitrous acid (as this 
decomposes the urea into carbon dioxide, water and 
nitrogen), or oxalic acid, added, to strongly acid reac- 

' tion. Upon cooling, urea nitrate or oxalate sepa- 
rates in white shining scales, or hexagonal tablets, 
the oxalate sometimes in four-sided prisms. (See 
Plate i, Fig. 2.) 

(b) Creatinin. C 4 H 7 N 3 0. Mix the greater portion 
of the alcoholic solution with a few drops of calcium 
hydrate, and then add calcium chloride as long as a 
precipitate is produced. Filter, reduce the filtrate 
on a water bath to 10 or 12 c.c, then pour this into 
a beaker, and after cooling, t add h c.c. of a pure 
alcoholic solution of zinc chloride. The precipitate 
collected after some hours' standing is examined 

G 



90 CHEMICAL ANALYSIS OF THE URINE. 

microscopically. (It usually forms delicate needles 
concentrically grouped, giving rise either to perfect 
rosettes or tufts.) 
(2) Hippuric acid. C 9 H 9 N0 3 . The two-thirds portion 
of the residue in III, feebly acidulated with hydro- 
chloric acid, is triturated with heavy spar powder 
(barium sulphate), and exhausted with alcohol. The 
alcoholic extract is saturated with sodium hydrate, 
the alcohol distilled off, and the syrupy liquid, after 
the addition of oxalic acid (to combine with the urea) 
evaporated to dryness on a water bath. Powder the 
residue and treat with ether, distilling off the latter 
and treating the warm residue to remove the excess 
of oxalic acid with calcium hydrate. Filter and re- 
duce the nitrate to a small volume and acidify with a 
little hydrochloric acid. After a short time hippuric 
acid will crystallize out and can be examined chemic- 
ally and microscopically. If the residue is gluey 
lactic acid is indicated. (See page 68.) If upon 
pouring some of the ethereal solution on water the 
well-known fat phenomena show upon the surface, fat 
is present. 
Hippuric acid crystallizes from a hot solution in delicate 
needles, from a cold saturated solution in milk-white, 
perfectly transparent four-sided prisms and columns, 
having two to four planes upon their extremities ; the 
principal form 'is a vertical prism (see Plate i, Fig. 3). 
(Distinction from benzoic acid, which crystallizes in 
tablets overlying each other.) On fusing, hippuric 
acid becomes first an oily fluid, and on cooling 
solidifies to a white crystalline mass, and this on being 
further heated to almost glowing, leaves a porous coke 



EXAMINATION OF URINE. 91 

in addition to sublimed benzoic acid and ammonium 
benzoate, and liberates an odor strongly similar to that 
of hydrocyanic acid. Strong nitric acid dropped into 
boiling hippuric acid, and evaporated to dryness 
leaves a residue, which, if heated in a small glass tube, 
sets free, like benzoic acid, an intense odor of oil of 
bitter almonds, from the formation of nitro-benzene. 
(3) The residue obtained in 1 is placed in a dish and 
dilute (1 part acid and 6 parts water) hydrochloric 
acid poured over it. The portion remaining undis- 
solved is collected on a small filter. 

(a) The earthy phosphates and other salts are found 
in the hydrochloric acid filtrate, and are precipi- 
tated by the addition of an excess of ammonium 
hydrate. 

(b) The residue on the filter contains mucin and 
uric acid. After washing with water, pierce the 
filter and with a stream of water from a wash 
bottle wash the residue into a small test tube, 
add 2 to 3 drops of sodium hydrate, warm and 
filter. 

(a) The undissolved residue is mucin. 

(/?) The filtrate will contain the uric acid, which 

yields crystals on being mixed with hydrochloric 

acid. 

IV. Urine coloring matters. (See page 15.) 

V. Glucose. (Test, page 55.~) 

VI. Hydrogen sulphide. (See page 73.) The urine 
smells of it, and colors paper saturated with lead 
acetate brown or black. 

VII. Inorganic substances. 

Evaporate 40-50 c.c. of urine to dryness on a water 



92 CHEMICAL ANALYSIS OF THE URINE. 

bath. Mix the residue with one to two grams of 
spongy platinum and ignite gently until all the 
carbon is burned off, when a greenish white mass 
remains. Reserve a small portion to test for iodine 
(see IX), the rest boil with water and obtain — 

A, a solution, and 

B, a residue. 

A. The solution is divided into four parts and examined 
for : — 

(1) Sulphuric acid. Acidify one part with hydro- 
chloric acid and add barium chloride; a white 
pulverulent precipitate, insoluble in acids. 

(2) Chlorine. Acidify a second portion with nitric 
acid and add silver nitrate ; a white curdy precipi- 
tate that blackens on exposure to light. 

(3) Phosphoric acid. Mix a third portion with sodium 
acetate, acetic acid, and add a few drops of ferric 
chloride; yellowish-white gelatinous precipitate. 
Another test is with ammonium molybdate, the 
liquid in presence of phosphoric acid is colored 
yellow and a yellow precipitate is produced. 

(4) Sodium. The remainder of the solution is evapor- 
ated to dryness and a small portion of the residue 
heated on a platinum wire in the inner flame 
of a blowpipe; yellow coloration imparted to 
flame. 

(5) Potassium. A portion of the residue in 4 is 
dissolved in a little water and platinum tetra- 
chloride added ; a yellow crystalline precipitate. 

B. The residue is extracted with hot hydrochloric acid, 
filtered, washed, and the filtrate examined for : — 



EXAMINATION OF URINE. 93 

(1) Iron. Heat a portion with a drop of nitric acid, 
and add sulphocyanide of potassium ; deep red 
coloration. 

(2) Calcium. Add an excess of sodium acetate to a 
second portion, and test with ammonium oxalate ; 
white precipitate. 

(3) Magnesium. Precipitate all the calcium as in 2; 
filter and add ammonium hydrate to filtrate ; there 
is formed a white precipitate of ammonium magne- 
sium phosphate. 

VIII. Ammonium salts. 50-100 c.c. of urine are mixed 
in a flask with sodium hydrate, and above it, by aid 
of the cork, a strip of moistened turmeric paper is 
hung. If ammonia is present, the paper rapidly be- 
comes brown, or if a glass rod moistened with hydro- 
chloric acid is held over the mouth of the flask, the 
well-known ammonium-chloride clouds are produced. 

IX. Iodine. Use the reserved portion of VII, put it in 
a porcelain crucible, moisten it with some drops of 
fuming nitric acid, and place a little starch paste on the 
under side of the lid of the crucible, which is then 
covered over the latter. In presence of iodine the 
starch is colored violet. The original urine contain- 
ing iodine can also be immediately distilled with sul- 
phuric acid. The latter method is, however, more 
complicated. 

H. Struve's colorimetric method for the estimation 
of iodine depends on a color scale, prepared by taking 
a potassium iodide solution of known strength, carbon 
bisulphide and a few drops of fuming nitric acid, and 
with this the color of the iodine solution from the 
urine is compared. 



94 CHEMICAL ANALYSIS OF THE URINE. 

Iodine can be estimated quantitatively by the fol- 
lowing method : 10-20 e.c. of palladious chloride, de- 
pending on the quantity of iodine in the urine, learned 
by previous qualitative tests, are heated in a corked 
flask, upon a water bath, and the urine containing 
iodine acidified with hydrochloric acid, and reduced 
by evaporation to a definite volume (10-20 c.c.) added, 
until all the palladium is precipitated as palladious 
iodide. Agitation of the mixture hastens the separa- 
tion. Small portions of the urine filtered off from 
time to time, and added to the urine under examina- 
tion, show when the reaction is complete. 

X. Kegarding the examination for the less important 
constituents of the urine, such as phenol, for the 
detection of which 20-30 kilos, of urine must be em- 
ployed ; further, benzoic and acetic acids, which 
only occur in decomposed urine; we can omit them, 
referring for details to larger works upon this subject. 
The same may be remarked of — 

XI. Butyric Acid, which is rarely found, and requires 
several kilos, of urine to detect it. 

XII. Inosite. (See page 66.) 

XIII. Allantoin. 

XIV. Xanthin. 

XV. Leucin and Tyrosin. (See page 70.) 

The quantitative determination of the various consti- 
tuents has already received treatment. 



URINARY CONCRETIONS. 95 

VII. 
URINARY CONCRETIONS. 

( Urinary Gravel and Calculi.) 

69. Concretions of the urine are deposits from the urine 
within the tracts (kidneys, ureter, bladder and urethra). 
Sometimes they are as small as grains of sand, and are con- 
sequently voided with the urine without much inconveni- 
ence. In such cases they are very abundant, and, as a 
rule, crystalline (urinary sand — gravel). Frequently they 
are larger, varying from the size of a pea to that of a small 
apple, and then cannot be voided (the true calculi). A 
sharp line of distinction between the two cannot be drawn ; 
generally, they are distinguished by difference in their 
form. 

70. The calculi consist mostly of a homogeneous mass, 
or of several concentric layers, frequently of chemically 
distinct substances, which have arranged themselves around 
a nucleus (very often a dried particle of mucin), and here 
gradually increased. 

71. We recognize and distinguish them readily under 
the microscope, especially where sand or particles have 
accidentally fallen into vessels and been mistaken for calculi 
by hypochondriacal patients. The grains, consisting mostly 
of silicates, are distinguished, by their appearance and 
physical deportment, from calculi, and rarely is a chemical 
examination necessary. 

Chemical Constituents of Calculi. 

72. They are essentially identical with those already 
mentioned under urinary sediments, and for their closer 



96 CHEMICAL ANALYSIS OF THE URINE. 

examination the student is referred to the preceding pages. 
Calculi may consist of: — 

(1) Uric acid and urates. 

(2) Xanthin. 

(3) Cystin. 

(4) Calcium oxalate. 

(5) Calcium carbonate. 

(6) Calcium phosphate. 

(7) Ammonium magnesium phosphate. 

(8) Proteid substances. 

(9) Urosteatite mixed with considerable quantities of 
silicic acid, aluminium oxide, etc. 

73. In making a chemical examination pursue the fol- 
lowing course : — 

After careful microscopic examinations (in calculi the 
different layers), which are important, because many cal- 
culi consist of but one of the above constitutents, pulverize 
the object to be examined, wash off with a little cold dis- 
tilled water, dry and ignite a sample upon platinum foil, 
over a Bunsen burner or spirit lamp. 

I. Either no or a very slight residue remains. 

II. The calculus appears to be incombustible or leaves 
a large residue after ignition. 

74. When there is no residue, or at most but a slight one, 
the following substances may be present : — 

. ' I burn without name. 

• Ammonium urate, J 

Xanthin, 

Cystin, 

Urosteatite, 

Proteid substances (as Fibrin, etc.), J 



burn with flame. 



URINARY CONCRETIONS. 97 

The chemical tests for these bodies are : — 

(1) Uric acid. Test by treating the powder with nitric 
acid and ammonium hydrate (murexide). Calculi of uric 
acid are relatively very frequent, and can attain a large 
size. Generally they are colored (yellow, reddish and red- 
dish-brown), rarely white, and possess usually a smooth 
surface and considerable hardness. 

(2) Ammonium urate. A portion of the sample treated 
with potassium or sodium hydrate liberates ammonia gas, 
recognized by white clouds formed about a glass rod, 
moistened with hydrochloric acid. 

Uric acid and ammonium urate are distinguished by the 
fact that uric acid is only slightly soluble in water, while 
ammonium urate dissolves much more readily, and in 
larger proportion. Calculi of ammonium urate are rare 
and generally of small size, of a clear (white or clay-yel- 
low) color, and rather earthy appearance. 

When the murexide reaction is not obtained the com- 
bustible concretion may consist of : — 

(3) Xanthin. Soluble in nitric acid without liberation 
of gas. In evaporating the solution there remains a resi- 
due of intense lemon yellow color, not reddened by am- 
monium hydrate, but soluble in sodium or potassium 
hydrate, with a deep reddish-yellow color. 

Guanin, has not yet been detected in urinary calculi. 
It yields a similar reaction to xanthin, therefore, care is 
necessary here. 

Calculi of xanthin are very rare, and thus far have been 
found in few instances. They have a clear brown (white 
to cinnamon-brown) color, are tolerably hard, with a waxy 
lustre, acquired by rubbing, and consist of concentric, 
easily soluble amorphous layers. 



98 CHEMICAL ANALYSIS OF THE URINE. 

(4) Cystin dissolves in ammonium hydrate, and crystal- 
lizes by spontaneous evaporation from this solution in very 
characteristic crystals, forming regular, six-sided tables, 
which occasionally are attached to a large six-sided rosette. 

On dissolving a calculus containing cystin in potassium 
hydrate, and boiling after the addition of a small quantity 
of lead acetate, there is formed a black precipitate of lead 
sulphide, which imparts to the mixture an inky tint. 

Cystin calculi are also very rare, of pale yellow color 
and smooth surface, with crystalline fracture and waxy, or 
greasy lustre. They are moderately soft, easily shaved, 
and the powder formed is much like that of soap. 

(5) Proteid substances do not exhibit the slightest trace 
of crystallization, diffuse, on burning, the odor of burning 
horn, insoluble in water, ether or alcohol, soluble in potas- 
sium hydrate, from which solution they are precipitated 
by acids. In acetic acid they expand and swell up, and 
are soluble in boiling nitric acid. 

Calculi from proteid substances (formed from fibrin and 
blood coagula) are very infrequent. 

(6) Urosteatite fuses when heated, without effervescence, 
swells up and liberates a very strong odor, recalling that 
of a mixture of shellac and benzene. It dissolves in potas- 
sium, or sodium hydrate, with saponification. Very solu- 
ble in ether. The residual urosteatite, after the evapora- 
tion of the ethereal solution, becomes violet on further 
warming. 

Calculi of this kind, like the preceding, are extremely 
rare. In the fresh condition they are soft, elastic, and 
resemble caoutchouc; on drying, they diminish in size, be- 
come brittle, light-brown to black, are moderately hard, 
and become softer on warming. 



URINARY CONCRETIONS. VM 

75. If the calculus did not burn, or left a large residue 
after ignition, it may consist of sodium, calcium, or mag- 
nesium urates, oxalate and carbonate of calcium, ammon- 
ium magnesium phosphate, and calcium phosphate. 

76. As we have already described the chemical tests of 
these substances, we will confine ourselves to only the most 
important points in what follows : — 

(1) Sodium, calcium, and magnesium urates do not 
readily occur as the sole constituents of calculi. Yet they 
sometimes are present in greater or less quantity in the 
calculus. To ascertain whether uric acid is united with 
such a base, boil the powder with distilled water, and filter 
while hot. The urates, more soluble in warm water than 
uric acid, pass into the filtrate. This is evaporated, ignited, 
and the bases in the residue tested for by the various pre- 
viously described methods. For the insoluble uric acid, 
see preceding pages. 

(2) Calcium oxalate blackens on ignition, by its con- 
version into calcium carbonate. Continued strong ignition 
leaves calcium oxide. Calculi of calcium oxalate are 
rather frequent, especially in children. They are either 
small, pale colored and smooth, Hemp-seed calculi, or 
they are larger, of rough exterior, bunchy, warty, 
colored dark brown on their surface and sometimes even 
black, Mulberry calculi. Owing to their rough surfaces, 
they irritate the urinary passages and induce serious dis- 
orders (bleeding, inflammation). 

(3) Calcium carbonate. Easily recognized by its effer- 
vescence with acids ; blackens also on ignition, resulting 
from organic substances present. 

(4) Ammonium magnesium phosphate and (basic) cal- 
cium phosphate occur generally intermixed with each 



100 



CHEMICAL ANALYSIS OF THE URINE. 



other ; do not burn on ignition, but fuse to a white, 
enamel-like mass ; hence called fusible calculi. After 
strong ignition they never react alkaline, differing in this 
respect from calculi of calcium oxalate and carbonate. 
In hydrochloric acid they dissolve without effervescence, 
and are re-precipitated from such an acid solution by 
ammonium hydrate. 

(5) In very rare cases calculi of neutral calcium phos- 
phate occur. In their chemical and physical properties 
they resemble the earthy phosphates, but differ from these 
in not containing any magnesium. 

COMPOSITION OF A SAMPLE OF URINE. 
Analysis by Miller. 



Water, 


956.37 




Urea, 


14.23] 

0.37 
. r , )P - j- 29.81 organic matter. 

0.16 J 


Uric acid, 
Extractive matter. . 


Mucus. 


Sodium chloride, 


7.22^ 




Phosphoric acid, 


2.12 




Sulphuric acid, 


1.70 




Calcium oxide, 


0.21 


' 13.82 inorganic matter 


Magnesium oxide, 


0.12 




Potassium oxide, 


1.92 




Sodium oxide, . 


0.53^ 





1000.00 43.63 total solids. 



L 



TABLE FOB Till". TENSION OF AQUEOUS VAl'oi; FOR 
TEMPERATURES FROM -2° TO 80°, CELSIUS, 

(BUNSEH . 



°c. 



Tension 

in 

Millimetre 



°0. 



Tension 
in 

Millinnfrs 



°C. 



Tend »n 

in 

Bfillimel re 



v. 



Tension 

in 
Rfillimet'n 



—2.0 


8.956 


6.2 


7.095 


—1.8 


4.01''. 


6.4 


7.198 


— 1.6 


4.078 


6.6 


7.292 


— 1.4 


4.140 


r>.* 


7. 192 


—1.2 


4.203 


7.0 


7.492 


—1.0 


4 267 


7.2 


7.596 


—0.8 


4.831 


7.4 


7. 699 


—0.6 


4.307 


7..; 


7.840 


—0.4 


4.463 


7.8 


7.010 


—0.2 


4.531 


8.0 


8.017 


0.0 


4 600 


8.2 


8.126 


-f0.2 


4 667 


8.4 


8.236 


0.4 


4.733 


8.6 


8.347 


0.6 


4.801 , 


8.8 


8.461 


0.8 


4.871 


9.0 


8.574 


1 


4.940 


9.2 


8.690 


1.2 


5.011 


9.4 


8.807 


1.4 


5.082 


9.6 


8.925 


1.6 


5.155 


9 8 


9.045 


1.8 


5 228 


i 10.0 


9. If, 5 


2.0 


5.302 


10.2 


9.288 


2 2 


5.378 


10.4 


9.412 


2.4 


.-, 45 1 


10.6 


9.587 


2.6 


5 530 


10.8 


9.665 


2.8 


5.608 


11.0 


9.792 


3.0 


5 687 


11.2 


9.02:1 


3.2 


5.767 


11.4 


10.054 1 


3.4 


5.848 


1 11.6 


10.187 


3.6 


5.930 


11.8 


10.322 


3.8 


6.014 


12.0 


10 457 


4.0 


6.097 


12.2 


10.596 


4.2 


6.183 


12.4 


10.734 


4.4 


6.270 


12.6 


10.875 


4.6 


6.350 


12.8 


11.019 


4.8 


6.415 


' 13.0 


11.162 i 


5.0 


6.534 


i 13.2 


11.309 


5.2 


6.625 


i 13.4 


11.456 


5.4 


6.717 


13.6 


11 605 


5.6 


6.810 


13.8 


11.757 


5.8 


6.904 


14.0 


11.908 


6.0 


6.998 | 


14.2 


12.064 i| 



14.4 
14.6 

1 1.s 
15 
1 5 2 
15.4 
15.6 
15.8 
16.0 
16.2 
16.4 
16.6 
16.8 
17.0 
17.2 
17.4 
17.6 
17.8 
18.0 
18.2 
18.4 
18.6 
1S.8 
19.0 
19.2 
19.4 
19.6 
19.8 
20.0 
20.2 
20.4 
20.6 
20.8 
21.0 
21.2 
21.4 
21.6 
21.8 
22.0 
22.2 
22.4 



12.220 
12.878 

12.58S 
12.699 
12.864 

13.029 
13.197 
18.366 
13.536 
18.710 
13.885 
14.062 
14.241 
11.421 
14.605 
14.7*90 
14.977 
15.167 
15.357 
15.552 
15.747 
15.945 
16.145 
16.346 
16.552 
16.758 
16.007 
17.179 
17.391 
17.608 
17.826 
18.047 
18.271 
18.495 
18.724 
18.954 
19.187 
19.423 
19.659 
10.001 
20.143 



22.6 
22.8 
23.0 
28.2 
23.4 
23.6 

24.0 
24.2 
24.4 
21.6 
24.8 
25.0 

25.4 
25.6 

25. S 
26.0 
26.2 
26.4 
26.6 
26.8 
27.0 
27.2 
27.4 
27.6 
27.8 
28.0 
28.2 
28.4 
28.6 
28.8 
29.0 
29.2 
29.4 
29.6 
29.8 
30.0 



20.389 
20.639 
20.888 

21.144 
21.400 
21.659 

21.921 
22.184 
22.158 
22.723 
22.096 
23.273 
23.550 
23.834 
24.119 
24.406 
24.697 
24.988 
25.288 
25.588 
25.891 
26.198 
26.505 
26.820 
27.136 
27.455 
27.778 
28.101 
28.438 
28.765 
29.101 
29.441 
20.782 
30.131 
80.470 
30.833 
31.190 
31.548 



ADDENDA. * 

Professor Wormley's paper (page 30) can be found in 
the American Journal of Medical Sciences, July, 1881, 
page 128. 



INDEX. 



Albumen ; 49 ; Galipe's test for, 52 ; 
Heller's test for, 52 ; in disease, 50 ; 
in presence of pus, 50 ; occurrence 
of, 51 ; qualitative detection of, 51 ; 
quantitative estimation of (Born- 

hardt), 52, 53; significance of 50 

Albuminuria 50 

Alkapton 12 

Allantom 12 

Ammonia, liberation from urea 12 

Ammoniacal salts, detection of 93 

Ammonium ; acid carbonate, 12 ; mag- 
nesium phosphate, 84, 86, 96; 

urate 84, 97 

Apparatus ; for the estimation of urea 
(Frontispiece), 29; for the estima- 
tion of urea (Htttfner), 27 ; for the 
fermentation of urine (Will and 
Varrentrapp\ 55 ; required in the 
examination of urine 14 

Barium ; chloride, standard solution 
of, 46 ; mixture for the removal of 

phosphates, etc 22 

Behavior of urine with chemical re- 
agents -. 10 

Benzoic acid 12, 13 

Biliary; acids, detection of, 70; col- 
oring matters (pigments), 12, 15, 

69; substances, detection of. 69, 70 

Bilirubin 70 

Biliverdin 16 

Blood ; in urine, 72 ; coloring matter 
(pigment) in urine, 81, 15, 71 ; cor- 
puscles 81 

Bright's kidney disease 50, 68, 86 

Butyric acid 68. 94 



Calcium; carbonate; 96; detection of, 
11, 93, 99 ; determination, 93; oxa- 
late, 78, 85, 96 ; in disease, 85 ; 
phosphate 78,79, 86, 96, 

Calculi ; 11, 95 ; combustible, 96 ; ex- 
amination of, 96 ; fusible, 100 ; non- 
combustible 

Cancerous masses 83 

Chemical; constituents of calculi....... 

Chlorides ; decrease of, 34 ; detection 
of, 10, 35 ; in urine, 34 ; occurrence 
of, 34 ; quantitative estimation 
(gravimetrically), 35 ; quantitative 
estimation (Liebig), 36 ; quantita- 
tive estimation (Neubauer & Mohr), 
37 ; quantitative estimation (Prim- 
bram), 40 ; quantitative estimation 
(Falck) 

Chlorine 

Cholicacid 



PAGK 

Composition of urine (Miller) 100 

Concretions, urinary 95 

Conferva? 77 

Creatin, normal urine constituent.... 10 
Creatinin ; 89 ; removal from urine, 
60, 61 ; solvent action upon cuprous 

oxide, 60; zinc chloride 60,89 

Cystin ; 12, 70, 79, 86, 96, 98 ; calculi, 98 
Cystinuria 79 

Diabetes mellitus 15 

Earthy phosphates ; 9, 10, 42, 91 ; es- 
timation of 45 

Epithelia 13 

Epithelial ; casts, 82 ; cells 82 

Fat; 12, 68, 78, 90; detection of 68 

Fehling's solution 62 

Fibrin 13, 71 

Frontispiece described 29 

Fungi 16, 83, 86 

Galactostasis 54 

Glucose; 12 ; fermentation 55 

Gravel, urinary 95 

Guanin 97 

Haematin 13, 72 

Hsematopyuria 51 

Hasmaturia 50 

Hemp-seed calculi 99 

Hippuric acid ; 10, 80, 90 ; in disease, 85 

Hyaline casts 82 

Hydrogen sulphide 13, 73, 91 

Hyposulphurous acid ; 34 ; estimation 
of , 34 

Indican; 9,74; estimation of 74 

Indigo 16 

Infusoria? 12, 86 

Inosite; 66,94; detection of 67 

Iodine; 93; estimation of. 93,94 

Iron 93 

Kidney; disease, Bright's, 50, 86; 
casts 82 

Lactic acid ; 10, 11 , 12, 68 ; detection of 68 
Leucin, 70,94; detection of 71 

Magnesium ; ammonium phosphate, 
96, 99 ; estimation of, 93 ; urate 99 

Maschke's modification of BOttger's 
test 57 

Mercuric nitrate, preparation of stan- 
dard solution 20 

Monada? 12 



103 



104 



INDEX. 



Mucin 13, 91 

Mucous coa^ula 80 

Mucus ; corpuscles, 80, 86 ; from the 

bladder in urine 11 

Mulberry calculi.. 99 

Murexide 32 

Mycodermas cerevisiae 11 



Nitrogen evolved from urea. 



30 



Oxaluria 85 

Oxaluric acid 33 

Oxymandelic acid 73 

Peptones in urine 51 

Phenol, or phenylic acid 10, 16, 94 

Phosphate of; sodium (standard solu- 
tion), 44; calcium, 78,79, 86; mag- 
nesium and ammonium 84, 86, 96 

Phosphates of the ; alkalies, 9, 42 ; al- 
kaline earths, 9, 10, 42 ; alkaline 

earths, estimation of... 45 

Phosphoric acid ; 41 ; detection of, 10, 
42, 92 ; estimation of, with uranium 

solution, 45; occurrence of 41 

Polarization ; apparatus, 66 ; of sugar 65 

Potassium 92 

Proteid substances ; 96, 98 ; removal 

from urine 57 

Pus 13, 81, 86 

Pyuria 51 

Qualitative and quantitative examin- 
ation of urine 87 

Reagents required in the examin- 
ation of urine 14 

Relations of sediments to the diagno- 
sis of disease 85 

Salkowski's observation upon Lie- 
big's urea method 25 

Salts, fixed, determination of 18 

Sarcina ventriculi Goodsir 84 

Seegur's observations on sugar 59 

Silica in urine 10 

Silver nitrate; solution, preparation of, 39 
Sodium ; acid urate, 78 ; detection of, 

92; urate 99 

Solid; residue of urine, 18; sub- 
stances in urine, proportion and 

estimation.... ,. 16,18 

Specific gravity of urine ; 10, 16 ; de- 
termination of 16 

Spermatozoids 13, 83, 86 

Substances affecting the reaction of 

sugar with reagents 61 

Succinic acid 12 

Sugar ; effect of minute quantities on 
Fehling's solution, 59; detection 
of, 54; according to Bence Jones, 
60; Blitz, 59; BOttger, 56; Campa- 
ni, 56; Fehling, 58; fermentation 
test, 55 ; indigo-carmine test, 55 ; 

N. B. — For uran. acetate, p. 43, 



PAGE 
I Knapp, 56 ; Mas^hke, 57 ; Moore, 
55; silver test, 55; Trommer, 58; 

quantitative estimation of. 61-65 

| Sulphates 10 

Sulphuric acid; 46; detection of, 10, 
92; gravimetric estimation, 46; 

volumetric estimation 46 

Sulpho-acids, estimation of 47 

Table for the tension of aqueous va- 
por 101 

Taurin 13, 69 

Taurocholic acid 69 

Torulacese 12 

Tubercular substances 87 

Tyrosin ; 12, 70, 77, 86, 94 ; tests for, 

71, 79, 80 

Triple phosphate 12,13,86 

Tube casts 86 

Uraemia 17,50 

Uranium ; phosphate, 42 ; solutions... 43 

Urates 11, 77, 85, 96, 99 

Urea; detection of, 19, 20, 89; errors 
in quantitative estimation of, 23; 
quantity of, 19; quantitative estim- 
ation (Fowler), 25; hypobromite, 
26 ; Liebig, 20, 22 ; Musculus, 31 ; 

recovery fr ...m urine 19 

Uric acid; action on Fehling's solu- 
tion, 60 ; as sediment, 11,. 78, 96 ; 
in disease or calculus, 31, 78, 85 ; 
occurrence of, 31 ; quantitative de- 
termination of, 32; recognition 

of 32,91,97 

Urinary; casts, 82; concretions, 9>; 
deposits, 11, 13, 75; sediments; 
amorphous, 76; crystalline, 76; de- 
tection of, under the microscope, 

76; organized 76 

Urine; 9, 10, 14, 15; color, 10; 
coloring matter; 10, 47, 91; esti- 
mation of, 48 ; concretions, 95 ; 
constituents ; abnormal, 49, 12; ac- 
cidental, 13 ; normal ; 10 ; quanti- 
ty of, 17, 18; fermentation; acid, 
11; alkaline, 11; gravel, 95; nor- 
mal, human, 9, 17 ; odor, 9, 16 ; of 
carnivorae (character), 9 ; of herbiv- 
orae (character), 9 ; physical char- 
acter of, 9 ; properties in diagnosis, 
14 ; reacti »n of, 10, 14 ; specific 

gravity of. 10 

Urobilin; 47; detection of, in urine, 48 

Urogl ncin 47 

Urophain; 47; test for 49 

Urosteitite 96, 98 

Uroxanthin 47 

Urrhodin 47 

Vibrionee 12,83 



Xanthin. 



10, 94, 96, 97, 



write-(C 2 H 3 2 ) 2 Ur0 2 + 3H 2 



